Micropropagation

Plants are conventially propagated either by sexual (through generation of seeds) or by asexual (by multiplication of vegetative parts) means. Emergence of modern methods of plant tissue culture has provided a very useful alternative which allows rapid propagation of desired plant species in a limited space under strict control of growth conditions. These plants are microbiologically sterile and thus exempt from quarantine regulations that may apply to export/import of normal plants. In vitro production of plants is generally described as micropropagation. Micropropagation (micro = small area of glass vessels; propagation = to increase the number of propagules) can be defined as asexual multiplication of plants in a small area of glass vessel (in vitro) under controlled physio-chemical conditions.


Micropropagation can be achieved by any one of the following pathways:

1. Stimulated axillary bud proliferation.

2. De novo adventitious shoot bud differentiation.

3. Somatic embryogenesis and

4. Callus organogenesis.


Every method has its own advantages and disadvantages and the choice of a particular pathway will depend upon them.


1. Stimulated shoot bud proliferation:- This method of in vitro propagation utilizes the clue from the basic physiological phenomenon in plants called as "apical dominance". Vascular plants with intermediate mode of growth have in their leaf axils subsidiary meristem with the potential for the growing into shoot. Apical dominance is the phenomenon by which the presence of apical bud causes a complete or partial inhibition of pre-existing lateral buds. In this method of micropropagation nodal segments having pre-existing but inhibited axillary shoot buds are used to stimulate proliferation of shoots in the presence of cytokinins. Nodal segments can be taken from both sexually mature or seedling (cotyledonary nodes) materials. In this method of micropropagation, the shoot multiplication may be initially slower than the other methods but with each passage the number of shoot increases logarithmically. This method is currently the most popular approach for clonal propagation of crop plants because the cells of the shoot apex are uniformly diploid and are least susceptible to genotypic changes under culture conditions.


2. De novo adventitious shoot bud proliferation:- Adventitious shoot buds are those which develop on any part of the plant such buds are not pre-existing and differentiate de novo. Bryophylium is the best example in nature where adventitious shoots buds differentiate on the margin of fleshy leaves. Such shoot buds develop roots and serve as propagules. Adventitious shoot buds can also be induced in vitro on a variety of explants such as hypocotyl segments, cotyledons, leaf, stem segments etc. Differentiated shoot buds on their subsequent subculture to fresh medium can be multiplied to obtain a regular crop of shoots.


3. Somatic embryogenesis:- Development of an embryo in nature is the consequence of fertilization. It is enclosed in cotyledons and the seed coat. Embryo is unique in the sense that it is bipolar in nature. On germination, it develops into shoot and root simultaneously. Such embryos which develop from somatic cells are called as somatic embryos. Somatic embryos can develop in two different ways:

(a). Direct somatic embryogenesis
(b). Indirect somatic embryogenesis

Direct somatic embryogeny can be induced on any explant such as hypocotyls, cotyledons, leaf, stem segments etc. Somatic embryo development begins with stimulation of few cells on the explant which from a globular structure subsequently developing into heart shaped, torpedo and cotyledonary stage embryos.The somatic embryos are borne naked and can easily be distinguished from the adventitious shoot buds. Somatic embryos are discrete units and lack vascular connection with the mother explant while adventitious shoot buds have continuity of vascularization with the mother explants.
In case of indirect somatic embryogeny, the embryo develops adventitiously with an intervening callus phase. Callus can be induced from a variety of explants and subsequently the embryos can be induced on it.




4. Callus organogenesis:- Callus is an unorganized mass of cells which can be induced on any explant grown on a suitable medium. Callus formation is basically a result of dedifferentiation (an explant which is differentiated tissue gets converted into dedifferentiated mass of cells). Callus can be multiplied in culture for indefinite period even in absence of light in culture room environment. Callus can again be induced to develop shoots by the phenomenon called redifferentiation (callus organogenesis). By manipulating plant growth regulator in the medium, both dedifferentiation (auxin rich medium) and redifferentiation (cytokinin rich medium) can be easily controlled. Durig callus formation, the cell to cell contact is lost and the cells seem to be in the state of "chaos".This result into somaclonal variation on account of genetic changes (ex. change in ploidy level) that occur in culture.

Arabidopsis Thaliana

Arabidopsis is an angiosperm, a dicot from the mustard family (Brassicaceae). It is popularly known as thale cress or mouse-ear cress. While is has no commercial value - in fact is considered a weed - it has proved to be an ideal organism for studying plant development.

Some of its advantages as a model organism:-

1. It has one of the smallest genomes in the plant kingdom: 115,409,949 base pairs of DNA distributed in 5 chromosomes (2n = 10).

2. Very little of this is "junk" DNA so most of the DNA encodes its 25,498 genes.

3. Transgenic plants can be made easily using Agrobacterium tumefaciens as the vector to introduce foreign genes.

4. The plant is small - a flat rosette of leaves from which grows a flower stalk 6-12 inches high.

5. It can be easily grown in the lab in a relatively small space.

6. Development is rapid. It only takes 5-6 weeks from seed germination to the production of a new crop of seeds.

7. It is a prolific producer of seeds (up to 10,000 per plant) making genetics studies easier.

8. Mutations can be easily generated (ex. by irradiating the seeds or treating them with mutagenic chemicals.

9. It is normally self-pollinated so recessive mutations quickly become homozygous and thus expressed.

10.Other members of its family cannot self-pollinate. They have an active system of self-incompatibility. Arabidopsis, however, has inactivating mutations in the genes - SRK and SCR - that prevent self-pollination in other members of the family.

11.However, Arabidopsis can easily be cross-pollinated to

   @ do genetic mapping and
   @ produce strains with multiple mutations.

Plant Hormone

Plant hormones also known as plant growth regulators (PGRs) and phytohormones) are chemicals that regulate a plant's growth. According to a standard animal definitions, hormones are signal molecules produced at specific locations, that occur in very low concentrations and cause altered processes in target cells at other locations. The word hormone is derived from Greek and means 'set in motion'. They are naturally produced within plants though very similar chemicals are produced by fungi and bacteria that can effect plant growth. The concentration of hormones required for plant responses are very low. Plant hormones affect gene expression and transcription levels, cellular division and growth.


The five major classes are:


1. Abscisic acid:- Abscisic acid also called ABA, was discovered and researched under two different names before its chemical properties were fully known,it was called dormin and abscicin 2. Once it was determined that the two latter named compounds were the same, it was named abscisic acid. The name "abscisic acid" was given because it was found in high concentrations in newly abscissed or freshly fallen leaves.

In plants that are water stressed, ABA plays a role in closing the stomata. Soon after plants are water stressed and the roots are deficient in water, a signal moves up to the leaves causing the formation on ABA precursors, these precursors moves to the roots which release ABA that is trans-located to the foliage through the vascular system, which regulates the potassium or sodium uptake within the guard cells then loses turgidity, closing the stomata.

2. Auxins:- Auxins are compounds that positively influence cell enlargement, bud formation and root initiation.They also promote the production of other hormones and in conjuction with cytokinins, they control the growth of stems, roots, flowers and fruits. Auxins were the first class of growth regulators discovered. They affect cell elongation by altering cell wall plasticity. Auxins decrease in light and increase where its dark. They stimulate cambium cells to divide and in stems cause secondary xylem to differentiate. Auxins act to inhibit the growth of buds lower down the stems, affecting a process called apical dominance and also promote lateral and adventitious root development and growth. Auxins especially NAA and IBA, are also commonly applied to stimulate root growth when taking cuttings of plants. The most common auxin found in plants is indoleacitic acid or IAA.

                                                           

3. Cytokinins:- Cytokinins are CKs are a group of chemicals that influence cell division and shoot formation. They were called Kinins in the past when the first cytokinins were isolated from yeast cells. They also help delay senescence or the aging of tissues are responsible for mediating auxin transport throughout the plant and affect internodel length and leaf growth. Cytokinins counter the apical dominance induced by auxins, they in

conjunction with ethylene promote abscission of leaves, flower parts and fruits.

4. Ethylene:- Ethylene is a gas that forms from the breakdown of methionine, which is in all cells. Ethylene has very limited solubility in water, and does not accumulate within the cell but diffuses out of the cell and escapes out of the plant. Its effectiveness as a plant hormone is dependent on its rate of escaping into the atmosphere. Ethylene is produced at a faster rate in rapidly growing and dividing cells especially in darkness. Ethylene affects cell growth and cell shape. Ethylene affects fruit ripening normally when the seeds are mature, ethylene production increases and builds-up within the fruit resulting in a climacteric event just before seed dispersal. The nuclear protein ETHYLENE INSENSITIVE 2 (EIN2) is regulated by ethylene production and in turn regulates other hormones including ABA and stress hormones.

5. Gibberellins:- Gibberellins or GAs include a large range of chemicals that are produced naturally within plants and by fungi. They were first discovered when japanese researches noticed a chemical produced by a fungus called Gibberella fujikuroi that produced abnormal growth in rice plants. They play a major role in seed germination, affecting enzyme production that mobilizes food production new cells need for growth. This is done by modulating chromosomal transcription. Gibberellins also reverse the inhibition of shoot growth and dormancy induced by ABA.

Java Programmes (Used In Bioinformatics)

 1.Addtion

public class Add
{
public static void main (String args[])
{

int x=20;
int y=30 ;
int x3=x+y;
System.out.println("Sum is"+x3);
}
}

out put:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Add ruchir amit
Sum is50
Finished executing

2.Addion (Command line argument)

public class Add1
{
public static void main (String args[])
{
String s1= args[0];
String s2= args[1];
int x1=Integer.parseInt(s1);
int x2=Integer.parseInt(s2);
int x3;
x3=x1+x2;
System.out.println("Sum is"+x3);
}
}
output:- 5 7
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Add1  5 7
Sum is12
Finished executing


3.WAP to check Prime no.

public class Prime
{
public static void main(String args[])
{
String s1=args[0];
int n=Integer.parseInt(s1);
int i;
for(i=2;i<n;i++)
{
 {if(n%i==0)
 break;
 }
 }
 if(i!=n)
 System.out.println("not prime");
 else
 System.out.println("prime");
   }
  }

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Prime 4
not prime
Finished executing

4.WAP to check armstrong no.

public class Armstrong
{
public static void main(String args[])
{
      int sum ,p,r;
      String s1= args[0];
      int y=Integer.parseInt(s1);
      r=y;
 sum=0;
while(y!=0)
{
 p=y%10;
 sum=sum+(p*p*p);
      y=y/10;
     }
     if (sum==r)
     {
     System.out.println("Armstrong");
     }
     else
     {
       System.out.println(" not Armstrong");
      }
    }
  }

output:-153
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Armstrong 153
Armstrong
Finished executing

5.WAP for heilstone series.

public class Heilstone
{
 public static void main(String args[])
 {
 String s1=args[0];
 int a=Integer.parseInt(s1);
 while(a>1)
 {
  if (a%2==0)
   a=a/2;
   else
   a=a*3+1;
   System.out.print(+a+",");
  }
  System.out.println(a);
 }
}
 output:12
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Heilstone 12
6,3,10,5,16,8,4,2,1,1
Finished executing


6.public class Series
{
 public static void main(String args[])
        {
         String s1=args[0];
         int n=Integer.parseInt(s1);
         int i,j;
         for(i=1;i<=n;i++)
            {
                for(j=1;j<=i;j++)
                {
                 System.out.print(j);
                }
                System.out.println(" ");
            }

        }

}

output:-5
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Series 5
1
12
123
1234
12345
Finished executing

7.public class Series2
{
 public static void main(String args[])
 {
  String s1=args[0];
  int n=Integer.parseInt(s1);
  int i,j;
  for(i=1;i<=n;i++)
  {
  for(j=1;j<=i;j++)
  {
  System.out.print(i);
  }
  System.out.println(" ");
  }
  }
 }

output:5
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Series2 5
1
22
333
4444
55555
Finished executing

8.public class Series3
{
 public static void main(String args[])
 {
 int i,j,k,s=30;
 for(i=1;i<=5;i++)
 {
 for(j=1;j<=s;j++)
 System.out.print(" ");
 for(k=1;k<=2*i-1;k++)
 {
 System.out.print("*");
 }
 System.out.println();
 s--;
 }
 }
 }
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Series3 5
                              *
                             ***
                            *****
                           *******
                          *********
Finished executing

7. WAP to print strigs according to asending order.
class stringorder
{
static String name[]={"RUCHIR", "NIKHIL", "SAUMYA", "ATUL", "AASTHA"};
public static void main(String args[ ])
{
int size=name.length;
String temp = null;
for (int i=0; i<size; i++)
{
for (int j=i+1 ;j<size;j++)
{
if (name[j].compareTo(name[i])<0)
{
temp= name[i];
name[i]=name[j];
name[j]=temp;
}
}
}
for (int i=0;i<size;i++)
{
System.out.println(name[i]);
}

output:C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" stringorder 5
AASTHA
ATUL
NIKHIL
RUCHIR
SAUMYA
Finished executing

8.WAP to print strigs according to asending order (no. of char).

public class Stringsort
{
public static void main(String args[])
{
int i,j;
String a[]={"anki","saumya","ruch","nikhi"};


for(i=0;i<a.length;i++)
{
for(j=0;j<i;j++)
{

if (a[i].length()>a[j].length())
{
String l=a[i];
a[i]=a[j];
a[j]=l;
}
}
}
for (i=0;i<a.length;i++)
{
System.out.println(a[i]);
}
}
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Stringsort 5
saumya
nikhi
ruch
anki
Finished executing

9. Default Constructor

class Student
{
int roll;
String nm;
Student()
{
roll=101;
nm="ruchir";
}
void display()
{
System.out.println("Name="+nm+"And roll="+roll);
}
}
public class Consdemo
{
public static void main(String args[])
{
Student t1=new Student();

t1.display();
}
}
output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Consdemo 5
Name=ruchirAnd roll=101
Finished executing

10. Parametrise Constructor

 class Student
{
int roll;
String nm;
Student(int r, String s)
{
roll=r;
nm=s;
}
void display()
{
System.out.println("Name="+nm+"And roll="+roll);
}
}
public class Conspara
{
public static void main(String args[])
{
Student t1=new Student(101,"Ruchir");

t1.display();
}
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Conspara 5
Name=RuchirAnd roll=101
Finished executing


11.Function Overloading
class area
{
final float pi=3.14f;
void calculate (int a)
{
int t=a*a;
System.out.println("Area of square="+t);
}


void calculate (int a,int b)
{
int t= a*b;
System.out.println("Area of reactangle="+t);
}

void calculate (float c)
{
float t= pi*c*c;
System.out.println("Area of circle="+t);
}
}
class function1
 {
 public static void main(String args[])
 {
 area a1=new area();
 a1.calculate(7);
 a1.calculate(4.2f);
 a1.calculate(8,3);
 }
 }

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" function1 5
Area of square=49
Area of circle=55.389595
Area of reactangle=24
Finished executing

12.Constructor overloading

class area
{
final float pi=3.14f;
void area (int a)
{
int t=a*a;
System.out.println("Area of square="+t);
}
area (int a,int b)
{
int t= a*b;
System.out.println("Area of reactangle="+t);
}
area (float c)
{
float t= pi*c*c;
System.out.println("Area of circle="+t);
}
}

class Functioncons
 {
 public static void main(String args[])
 {
 area a1 = new area(7);
 area a2 = new area(7,8);
 area a3 = new area(5.2f);
}
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Functioncons 5
Area of circle=153.86002
Area of reactangle=56
Area of circle=84.905594
Finished executing

13.Single inheritance
class item
{
int pr,qty;
void getitem(int a,int b)
{
pr=a;
qty=b;
}
void display()
{
System.out.println( "price="+pr+"and quality="+qty);
}
}
class total extends item
{
int total;
void calculate()
{
total=pr*qty;
System.out.print("total value="+total);
}
}
class inherit
{
public static void main(String args[])
{
total t1= new total();
t1.getitem(6,25);
t1.display();
t1.calculate();
}
}

output:
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" inherit 5
price=6and quality=25
total value=150Finished executing

14.Interface
interface Area
{
final float pi = 3.14f;
void calculate(int a);
}
class Circle implements Area
{
  public void calculate(int a)
{
float t=a*a*pi;
    System.out.println("Area of circle="+t);
}
}
class Square implements Area
{
public void calculate(int a)
{
int t=a*a;
System.out.println("Area of square="+t);
}
}
public class Interfacedemo
{
public static void main(String args[])
{
Area a1;
Circle c1=new Circle ();
  Square s1=new Square ();
a1=c1;
a1.calculate(6);
a1=s1;
a1.calculate(4);
}
}
 output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" inherit 5
price=6 and quality=25
total value=150Finished executing


15. Multilevel Inheritance

class item
{
String na;
int pr,qty;
item (int p,int q,String s)
{
na=s;
pr=p;
qty=q;
}
void display()
{
System.out.println("name="+na + "price="+ pr + "quality="+qty);
}
}
class user extends item
{
int to_user,to_price;
int no;
user ( String a,int b,int c,int d)
{
super(c,d,a);
no=b;
}
void display()
{
super.display();
to_price=no*pr*qty;
to_user=qty;
System.out.print("total price="+to_price+ "and total user="+to_user );
}
}

 class ruc
{
public static void main(String args[])
{
user a1=new user( "pen",10,105,25);
a1.display();
}
}
 output:-
 C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" ruc 5
name=penprice=105quality=25
total price=26250and total user=25Finished executing

16.Multilevel and Multiple Inheritance

class student
{
int roll;
String nm;
void getdata(int r,String s)
{
roll=r;
nm=s;
}
void dispdata()
{
System.out.println("name="+nm+ "roll no.="+roll);
}
}
class marks extends student
{
int m1,m2;
void getmarks(int a,int b)
{
m1=a;
m2=b;
}

void dispmarks()
{
System.out.print("sub_1="+m1+ "and sub_2="+m2 );
}
}
interface bonus
{
int b=20;
void dispbonus();
}
class result extends marks implements bonus
{
int tot;
public void dispbonus()
{
System.out.print("bonus marks="+b);
}
void dispall()
{
dispdata();
dispmarks();
dispbonus();
tot=m1+m2+b;
System.out.print("Total marks="+tot);
}
}
class ruch
{
public static void main(String args[])
{
result r1=new result();
r1.getdata(101,"ruchir");
r1.getmarks(50,60);
r1.dispall();
}
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" ruch 5
name=ruchirroll no.=101
sub_1=50and sub_2=60bonus marks=20Total marks=130Finished executing


17. Multiple Inheritance
class A extends Thread
{
public void run()
{
int i;
for (i=1 ;i<=5;i++)
{
System.out.println("from thread A="+i);
}
System.out.println("exit from thread A");
}
}

class B extends Thread
{
public void run()
{
int j;
for (j=1 ;j<=5;j++)
{
System.out.println("from thread B="+j);
}
System.out.println("exit from thread B");
}
}

class c extends Thread
{
public void run()
{
int k;
for (k=1 ;k<=5;k++)
{
System.out.println("from thread C="+k);
}
System.out.println("exit from thread C");
}
}
class prioritythread
{
public static void main(String args[])
{
A t1=new A();
B t2=new B();
c t3=new c();
t1.start();
t2.start();
t3.start();
System.out.println("exiting from main");
}
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" prioritythread 5
exiting from main
from thread B=1
from thread B=2
from thread B=3
from thread B=4
from thread B=5
exit from thread B
from thread A=1
from thread A=2
from thread A=3
from thread A=4
from thread A=5
exit from thread A
from thread C=1
from thread C=2
from thread C=3
from thread C=4
from thread C=5
exit from thread C
Finished executing

18. Multiple Inheritance with exceptional handleing

class A extends Thread
{
public void run()
{
int i;
for (i=1 ;i<=5;i++)
{
System.out.println("from thread A="+i);
if(i==2)
yield();
}
System.out.println("exit from thread A");
}
}

class B extends Thread
{
public void run()
{
int j;
for (j=1 ;j<=5;j++)
{
System.out.println("from thread B="+j);
}
System.out.println("exit from thread B");
}
}

class c extends Thread
{
public void run()
{
int k;
for (k=1 ;k<=5;k++)
{
System.out.println("from thread c="+k);
if (k==2)
try
{ Thread.sleep(10);}
catch (Exception e1)
{}
}
System.out.println("exit from thread C");
}
}

class thread2
{
public static void main(String args[])
{
A t1=new A();
B t2=new B();
c t3=new c();
t1.start();
t2.start();
t3.start();
System.out.println("exiting from main");
}
}

Output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" thread2 5
from thread A=1
from thread A=2
exiting from main
from thread c=1
from thread c=2
from thread A=3
from thread A=4
from thread B=1
from thread A=5
from thread B=2
exit from thread A
from thread B=3
from thread B=4
from thread B=5
exit from thread B
from thread c=3
from thread c=4
from thread c=5
exit from thread C
Finished executing

19.Thread priority

class A extends Thread
{
public void run()
{
int i;
for (i=1 ;i<=5;i++)
{
System.out.println("from thread A="+i);
}

System.out.println("exit from thread A");
}
}
class B extends Thread
{
public void run()
{
int j;
for (j=1 ;j<=5;j++)
{
System.out.println("from thread B="+j);
}
System.out.println("exit from thread B");
}
}
class c extends Thread
{
public void run()
{
int k;
for (k=1 ;k<=5;k++)
{
System.out.println("from thread C="+k);
}
System.out.println("exit from thread C");
}
}
class priority_th
{
public static void main(String args[])
{
A t1=new A();
B t2=new B();
c t3=new c();
t1.setPriority(Thread.MIN_PRIORITY);
t3.setPriority(Thread.MAX_PRIORITY);
t1.start();
t2.start();
t3.start();
System.out.println("exiting from main");
}
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" priority_th 5
exiting from main
from thread C=1
from thread C=2
from thread C=3
from thread B=1
from thread C=4
from thread A=1
from thread C=5
from thread B=2
exit from thread C
from thread A=2
from thread A=3
from thread A=4
from thread B=3
from thread A=5
from thread B=4
exit from thread A
from thread B=5
exit from thread B
Finished executing

20 runnable interface:-

class A extends Thread
{
public void run()
{
int i;
for (i=1 ;i<=5;i++)
{
System.out.println("from thread A="+i);
}

System.out.println("exit from thread A");
}
}
class B extends Thread
{
public void run()
{
int j;
for (j=1 ;j<=5;j++)
{
System.out.println("from thread B="+j);
}
System.out.println("exit from thread B");
}
}
class c extends Thread
{
public void run()
{
int k;
for (k=1 ;k<=5;k++)
{
System.out.println("from thread C="+k);
}
System.out.println("exit from thread C");
}
}
class runnableinter
{
public static void main(String args[])
{
A t1=new A();
B t2=new B();
c t3=new c();
Thread a1=new Thread(t1);
Thread a2=new Thread(t2);
Thread a3=new Thread(t3);
a1.start();
a2.start();
a3.start();
System.out.println("exiting from main");
}
}

output:
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" runnableinter 5
from thread A=1
from thread B=1
from thread B=2
from thread B=3
exiting from main
from thread B=4
from thread B=5
exit from thread B
from thread C=1
from thread A=2
from thread C=2
from thread A=3
from thread C=3
from thread A=4
from thread C=4
from thread A=5
from thread C=5
exit from thread A
exit from thread C
Finished executing

21.Synchronization:-

class One
{
      synchronized void display(int num)
      {
            System.out.println(" " + num);
            try
            {
               Thread.sleep(1000);

            }
            catch(InterruptedException e)
            {
                System.out.println("Interrupted");
            }
            System.out.println("Exiting");
        }
}

class Two implements Runnable
{
      int n;
      One o1;
      Thread t;
      public Two(One a1,int t1)
      {
            n=t1;
            o1=a1;
            t=new Thread(this);
            t.start();
      }
      public void run()
      {
             o1.display(n);
      }
}

class Synchronize
{
      public static void main(String args[])
      {
             One t1=new One();
             int digit=10;
             Two t2=new Two(t1,digit++);
             Two t3=new Two(t1,digit++);
             Two t4=new Two(t1,digit++);
             try
             {
                t2.t.join();
                t3.t.join();
                t4.t.join();
             }
             catch(InterruptedException e)
             {
             }
       }
}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Synchronize 5
 10
Exiting
 12
Exiting
 11
Exiting
Finished executing

22.Daemon (type of thread)
class Daemon implements Runnable
{
      static Thread t1,t2;
      public Daemon()
      {
             t1=new Thread(this);
             //t1.start();
             t2=new Thread(this);
             t2.setDaemon(true);
             t1.setDaemon(true);
      }
      public void run()
      {
             System.out.println("Number of threads running " +Thread.activeCount());
             System.out.println(t1.isDaemon());
             System.out.println(t2.isDaemon());
      }
             public static void main(String args[])
             {
                    new Daemon();
                    System.out.println("Number of threads running " +Thread.activeCount());
                    System.out.println(t1.isDaemon());
                    System.out.println(t2.isDaemon());
             }

}

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Daemon 5
Number of threads running 1
true
true
Finished executing

23. Static class
 class abc
{
void display()
{
System.out.println("non static mathod");
}
static void hello()
{
System.out.println("static mathod");
}
}

 class staticdemo
 {
 public static void main(String args[])
 {
 abc t1 = new abc();
 t1.display();
 abc.hello();
 }
 }

output:-
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" staticdemo 5
non static mathod
static mathod
Finished executing

24.Exceptional handling
class Exceptiondemo
{
public static void main(String args[])
{
try
{
String s=args[0];
int a= Integer.parseInt(s);
int c=a*a;
System.out.println(c);
}
catch (ArrayIndexOutOfBoundsException e1)
{
System.out.println("No arument passed");
}
catch(NumberFormatException e2)
{
System.out.println("Invalid type");
}
catch (Exception e3)
{
System.out.println(e3);
}
System.out.println("Program ending");
}
}

output 1:-5
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Exceptiondemo 5
25
Program ending
Finished executing
output 2:-5.2
C:\Program Files\Java\jdk1.6.0_03\bin\java.exe  -classpath "C:\Program Files\Java\jdk1.6.0_03\jre\lib\rt.jar;C:\Program Files\Java\jdk1.6.0_03\lib\tools.jar;E:\java_programs" Exceptiondemo 5.3
Invalid type
Program ending
Finished executing

25. Different with help of Applet :-
import java.applet.Applet;
import java.awt.*;
public class Red extends Applet
{
public void paint(Graphics g)
{
g.drawRect(50,50,150,400);
g.drawOval(100,110,50,50);
g.setColor(Color.red);
g.fillOval(100,110,50,50);
g.drawOval(100,190,50,50);
g.setColor(Color.yellow);
g.fillOval(100,190,50,50);
g.drawOval(100,270,50,50);
g.setColor(Color.green);
g.fillOval(100,270,50,50);
}
}

26. Applet (text box ,click box)

import java.awt.*;
import java.awt.event.*;
import java.applet.Applet;
public class Appdemo extends Applet implements ActionListener
{  TextField t1,t2,t3;
    Button b1,b2,b3,b4;
 public void init()
 {
 Label l1=new Label ("enter a no.");
 Label l2=new Label ("Another no");
 Label l3=new Label("Answer");
t1=new TextField(20);
t2=new TextField(20);
 t3=new TextField ("30");
  b1=new Button("Add");
  b2=new Button("Sub");
  b3=new Button("multi");
  b4=new Button("div");
 add(l1);
 add(t1);
add(l2);
add(t2);
add(l3);
add(t3);
add(b1);
add(b2);
add(b3);
add(b4);
b1.addActionListener(this);
b2.addActionListener(this);
b3.addActionListener(this);
b4.addActionListener(this);
}

public void actionPerformed(ActionEvent e1)
{
String s1,s2,s3;
int n1,n2,n3;
s1=t1.getText();
s2=t2.getText();
n1=Integer.parseInt(s1);
n2=Integer.parseInt(s2);
if(e1.getSource()==b1)
{
n3=n1+n2;
}
else if(e1.getSource()==b2)
{
n3=n1-n2;
}
else if(e1.getSource()==b3)
{
n3=n1*n2;
}
else if(e1.getSource()==b4)
{
n3=n1/n2;
}
s3=String.valueOf(t3);
t3.setText(s3);
}
}

HTML page:-Appdemo.html
<HTML>
<HEAD>
    <TITLE></TITLE>
</HEAD>
<BODY>
 <Applet code="Appdemo.class" width ="600" height="800">
 </Applet>
</BODY>
</HTML>

27. traffic light with help of Applet

import java.applet.Applet;
import java.awt.*;
public class Traffic extends Applet
{
 public void init ()
  {
   setBackground(Color.white);
   }
 public void paint (Graphics g)
  {
   g.setColor(Color.black);
   g.fillRect(0,0,50,90);
   g.setColor(Color.red);
   g.fillOval(10,20,20,20);
   g.setColor(Color.yellow);
   g.fillOval(10,40,20,20);
   g.setColor(Color.green);
   g.fillOval(10,60,20,20);
   }
}

html page:- traffic.html
<HTML>
<HEAD>
    <TITLE></TITLE>
</HEAD>
<BODY>
<Applet Code ="Traffic.class" width ="300">
</Applet>
</BODY>
</HTML>

2.traffic light (blinking)

import java.applet.Applet;
import java.awt.*;
public class Trafficlight extends Applet
{
 public void init ()
  {
   setBackground(Color.white);
   }
 public void paint (Graphics g)
  {
   g.setColor(Color.black);
   g.drawRect(0,0,50,90);
   for( ; ;)
   {
   g.setColor(Color.red);
   g.fillOval(10,20,20,20);
   g.setColor(Color.black);
   g.fillOval(10,40,20,20);
   g.fillOval(10,60,20,20);
   try
   { Thread.sleep(2000);}
   catch(Exception e)
   {}
   g.fillOval(10,20,20,20);
   g.setColor(Color.yellow);
   g.fillOval(10,40,20,20);
   g.setColor(Color.black);
   g.fillOval(10,60,20,20);
   try
   { Thread.sleep(2000);}
   catch(Exception e)
   {}
   g.setColor(Color.black);
   g.fillOval(10,20,20,20);
   g.fillOval(10,40,20,20);
   g.setColor(Color.green);
   g.fillOval(10,60,20,20);
   try
   { Thread.sleep(2000);}
   catch(Exception e)
   {}
   }
  }
}
 html page:-traffic.html
<HTML>
<HEAD>
    <TITLE></TITLE>
</HEAD>
<BODY>
<Applet Code ="Trafficlight.class" width ="300">
</Applet>
</BODY>
</HTML>

Plant Cell Growth

Growth is the self-multiplication of living material, the protoplasm it self. Growth is an increase in size (volume or length) duw to cell divisions and subsequent enlargement. It is an increase in dry weight or bulk of an organism associated with development.


Development is defined as an ordered change or progress, often towards a higher, more ordered or more complex state. Development may take place with growth and growth may take place with development.



Population growth is the increase in the total mass of cells making the population, or the increase in the total number of cells of the population. Individual growth is the increase in mass of the individual cell.

WHY GROWTH OCCURS?



Growth is expressed as the division of a cell to form two cells and the enlargement of the newly divided cells. When we say that cells double in all their constituents and then divide in half, we are obviously describing the average case. When we observe individual cells, however, we find deviations from the average. In some instances, the growth rate is constant rather than accelerating.

Mass of cells doubles and halves only in an average way and not in an exact way. Cells divide when they are ready and they are ready only when they have completed certain preparations for division.

CRITICAL MASS THEORY:-


In 1908 the German biologist Richard von Hertwig proposed the theory of the critical mass. In Hertwig's view a growing cell eventually reaches a size at which the ratio between the masses represented by cytoplasm on the one hand and the cell nucleus on the other becomes limiting. When that limiting ratio is reached, he proposed, some instability sets which triggers cell division. The nucleus-cytoplasm relation theory, as hertwig called it, does not hold up very well when mass alone is considered to be the factor that triggers cell division. Many exceptions to this hypothesis have accumulated that, although the division of cells usually parallels, an increase in cytoplasmic mass, this increase alone is not generally accepted as the fundamental mechanism
controlling cell cycle.

AGE OF CELLS:-



Cells taken from old cultures are often referred to as old cells, The age of a single cell cannot be greater than the time between two divisions. A cell, when first formed by cell division, although often called a young cell, contains materials both genetic and protoplasmic, which pre-existed in the parent cell. Age of the cell depend on its life spam. The life spam begins when the daughter cell is completely separated from the parental cell, this is called inception. It ends when this daughter cell itself divide later to give new daughter cells, this is known as termination. The period between inception and termination is called the generation time. This is actually the visible generation time that is observed. We can define young population of cells as the one, which is still actively growing and contains fewer cells that it is possible to obtain from that culture under tha same conditions, but old population is defined as that one which has reached the maximum number of cells under similar conditions. There are three types of following generation times :-

1. The visible generation time which is the period between inception and termination. It depends on division of cell wall.

2. Nuclear generation time depends on division of the nucleus. It starts from the moment the nucleus divides to the complete ability to divide again in the new daughter cells.

Generation time is given the symbol t.

SYNCHRONOUS AND SYNCHRONIZED GROWTH:-

Synchronous growth cultures are produced without metabolic shock. But synchronized culture are two ones in which we produce step-wise growth, the synchrony is induced by physiological shock. The principal means of doing synchronous growth are as follows :

1. Separation by physical means of cells in the population that are all at the same stage of the cellular life   cycle. Ex. by filteration, centrifugation etc.

2. Attachment of growing cells to a membrane, from which one product of cell division is shed. i.e., age selection. 

BALANCED AND UNBALANCED GROWTH:-

Exponential growth, whether in batch or continuous cultures, is the balanced growth. That is all cellular constituents are manufactured at constant rates relative to each other. If nutrient levels or other environmental conditions change, unbalanced growth results because the rates of synthesis of cell components vary relative to one another until a new balanced stage is reached.

Unbalanced growth occurs when a bacterial population is shifted down from a rich medium to a poor one. The organism may previously have been able to obtain many cell components directly from the medium. When shifted to a nutritionally inadequate medium, they need time to make the enzyme required for the biosynthesis of unavailable nutrients. Consequently cell division and DNA replication continue after the shift-down, but net protein and RNA synthesis slow.

Computer Aided Drug Design

Tissue Culture Techniques To Increase Genetic Variability

The haploid plantlets (n chromosomes) are treated with colchicine to produce fertilie homozygous lines, called double haploid lines. The haplodiplodisation technique could give immediately new elite genotypes, hybrid parents after in vitro propagation, as asparagus supermales (MM) or useful genetic material to establish gene mapping.

PROTOPLAST CULTURE & MICROPROPAGATION :



Protoplasts are the smallest unit able to regenerate a whole plant. Therefore protoplats cultures can serve to enlarge genetic variability by introducing somaclonal variations. However, the main interest of protoplasts, is their capacity to fuse and to produce hybrids or cybrids. Naked protoplasts can accept without rejection external elements: nuclei, cytoplasmic organelles, liposomes, containing genetic information.

The hybridization programme is hampered in some cases because of sexual incompatibility, Synthetic production of hybrids has, however becomes possible through the novel technique of protoplast culture and their fusion. The isolation culture, and fusion of protoplast are one of the most fascinating fields of research, through still in developing stage. The protoplast culture technique can be suitably used for microinjection and other genetic engineering experiments. The technique are important, specially because of their far-reaching effects on crop improvement by somatic hybridization and cell modification. The protoplast culture can be regenerated into an entire plant. The discovery of enzymes which could separate the cells for isolating protoplasts and exploring the possibilities of genetic engineering.

SOMATIC HYBRIDS & CYBRIDS :

The first protoplast fusion application was a cytoplasmic transfer from one genotype to another to induce male sterility (CMS) from mitochondrial origin. These male sterile hybrids are interesting to produce F1 hybrids (Brassica, Cichorium).

Since long protoplast fusion was proposed as a novel and important method for producing hybrid plants that can be obtained by sexual means. Mechanical or enzymatic removal of cell wall from a plant cell yields, a protoplast, which is bound by plasma membrane. An isolated protoplast, under suitable culture condition can regenerate entire plant. Plasmolyzing cell prior to enzymatic treatment helps to facilitate protoplast isolation. The isolated protoplast, having lost the protective cell wall, has to be protected against the osmotic shock by keeping them in the isotonic state. 13% mannitol is a good agent in this context. A variety of enzymes are available for protoplast isolation. Most common are cellulose, pectolyase and macerozyme. The viable protoplast tend to synthesize new cell wall within few hours to few days of culture divide and re-divide, produce clumps of cells which finally produce plantlets.

PRODUCTION OF VIRUS FREE PLANTS :

In 1952, Morel and Martin were successful in regenerating a virus-free dahlia plant by the excision of some meristematic domes from virus infected shoots. Semal and Lepoivre (1992) reported that a virus-free sweet potato was producing 40T/ha in china by comparison of the 20 T/ha produced before meristem culture.

Virus eradiction is dependent on several parameters. But to take advantage of the non uniform and imperfect virus distribution in the host plant body, the size of the excised meristem should be as small as possible. For Stone (1963), only tips between 0.2 and 0.5 mm most frequently produce virus free carnation plants. The explants smaller than 0.2 mm can't survive and those larger than 0.7 produce plants that still contain mottle virus.

There are various explanations have been given: absence of plasmodesm in the meristematic domes, competition between synthesis of nucleoproteins for cellular division and viral replication, inhibitor substances, absence of enzymes present only in the cells of the meristematic zones and suppression by excision of small meristematic domes. This last proposal could explain why some potato plant showing virus particles in the meristematic domes, could regenerate a virus free plant.

SHOOT TIP MICROGRAFTING :

When meristematic tip culture fails, it is possible to graft small meristematic domes on young seedlings growing in vitro. In this way, Navarro et al. (1975) eradicted all the virus diseases from spanish Citrus orchards. This technique was also very successful in eliminating virus diseases from peach tress (Mosella et al 1980).

MICROPROPAGATION TECHNIQUES:

During the micropropagation process, the genetic stability of new shoots dependent upon their origin. Axillary shoots issue from pre-existing buds and are normally true to type, indeed the meristematic cells are genetically very stable. Adventitious shoots, such as somatic embryos, are neoformed buds developed directly on some organs, or indirectly through a callus phase formed on this organ. So, if the mother plant presents a cell mosaic or chimaeric tissues, risks of genetic variation exists. It is similar in the case of an indirect regeneration, when the callus phase is too long.

PROPAGATION BY AXILLARY SHOOTING :

This technique has proved to be the most applicable and reliable method of in vitro propagation. Axillary shoot growth is stimulated by overcoming apical meristem dominance. Commercial tissue culture laboratories are now able to propagate a large number of herbaceous ornamental species and several woody plants in this way. However, the propagation of Pelargonium, Howea and a few other horicultural plants are always difficult to propogate by axillary branching.

PROPAGATION BY DIRECT OR INDIRECT ORGANOGENESIS :

Adventitious shoots could arise directly from the tissue of explants without callus formation. Several plants of tha family gesneriaceae (Saintpaulia, Streptocarpus) regenerate directly buds on leaf explants, likewise Lilium regenerate on scales. However, more often, like for Ficus lyrata, adventitious buds appear on callus. While coffee, cocoa trees and many conifers are produced by somatic embryogenesis developed on callus or cell suspensions.

IMPROVEMENT OF AXILLARY BRANCHING :

The cost of micropropagated plantlets is also an important limitation of the techniques. In New zealend, where they produce 2-3 million micropropagated radiata pine per annum, the relative cost of micropropagated planting stock had dropped from 13,8 times the cost of seedlings in 1988 to 6,9 by 1993 (Smith, 1997). To reduce manpower costs, several improvements have been proposed.. The more simple method was in vitro layering developed by Wang(1977) to clone PVX-free potato plants.The first plantlets placed on the medium in a horizontal position developed axillary shoots. They are harvested by cutting one centimeter above the medium surface, at 3 weeks intervals. A similar technique called "hedging system" by Aitken christie and Jones (1987) was later used to produce Pinus radiata. Since 1988, Duhem was producing vary large quantities of Eucalyptus plantlets in petri dishes without anti-gibberellin but in complete darkness. Transfers from one petri to another is made by a simple squashing.

SOMATIC EMBRYOGENESIS PROPAGATION :

For genetically stable species, somatic embryogenesis offers a very fast scaling-up system, especially when it's possible to produce embryos in bioreactors. Only a few model plants are successfully produce by such technology: carrot, celery. Other applications remian at the experimental stage: coffee, oil, palms, conifers, Euphorbia pulcherrima and several other horticultural species. 

Several bottlenecks limit the use of this interesting technology. one of main problems is genetic stability. Therefore, despite the clonal nature of nucellar embryos, different morphological anomalies can occur among mango somatic embryos, as it was also observed in Citrus plants derived from nucellar cultures (Litz et al 1993). Another difficulty is the loss of embryogenic capacity overtime, a phenomenon observed with different species. It is also important that somatic embryogenic lines of conifers are always originated from immature embryos.

SYNSEEDS :

Another very interesting possibility of the somatic embryogenesis technology has been developed during the past 15 years by Redenbaugh and his team (1991, 1993). They were able to encapsulate somatic embryos by hydrogel coatings (sodium alginate), producing single embryo artificial seeds. To date, some improvements offer the possibility to directly plant the artificial seeds in the greenhouse on special substrates (vermiculite, sans). This methodology will provide in future a good technique to reduce the cost of transplants. 

SOMACLONAL VARIATIONS :

The production of plantlets by callus regeneration, cell suspensions, protoplast cultures could present some deviations with regard to the mother plant. This is the way to increase the genetic variability. Associated with a selective pressure (Stress to toxins, pH, salinity, cold). it's used to obtain resistant lines.Indeed after regeneration, plants can express new potentialities rarely obtained another way. Stable and profitable variants are selected and introduced in breeding programmes. In 1976, a pelargonium cv Velvet Rose was created by this technique (Reisch, 1983).

Powerpoint Presentation On Analog Based Drug Design

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Microbial Biopesticides, Biofungicide, Herbicides And Agricultural Antibiotics

Several biopesticides are in use today. Biopesticide products are based on natural agents such as microorganisms and fatty acid compounds. They are toxic to targeted pests (such as the European corn borer) and do not harm humans, animals, fish, birds or beneficial resistance to conventional pesticides. One of the most common microorganisms used in biologically based pesticides is the Bacillus thuringiensis, or Bt, bacterium. Several of the proteins the Bt bacterium products are lethal to individual species of insects. Using Bt bacteria in pesticide formulations can eliminate target insects without relying on chemically based pesticides. It is also possible to use pheromones in pest control. Pheromones are naturally occuring substances that insects produce to attact mates. In pest control, pheromones are used to attract insects away from crop plants. For example, pheromone-based traps were used to control fruit fly infestations in california. The European corn borer, one of the most prevelant pests, costs the united states $1.2 billion crop damage each year.

Using biotechnology, it is possible to make crop plants tolerant of specific herbicides. When the herbicide is sprayed, it will kill the weeds but have no effect on the crop plants. This lets farmers reduce the number of times herbicides have to be applied and reduces the cost of producing crops and damage to the environment.

The contributions of biotechnology to the practice of medicine through the discovery and development of a number of potent antibacterial antibiotics is widely recognized. A major effort concerned with the discovery of new structures with antibacterial activities and the chemical modification of existing natural products is still ongoing. Although these antibiotics have found widespread use as growth promoters in the animal health area, only more recently were microbial products produced primarily for animal growth or agricultural purposes, for instance monensin as a coccidiostat for poultry and a growth permittant for ruminant animals. Bacteria, fungi and virus are the most commonly researched as potential microbial biocides. The development of low cost production methods and successful formulation of products such as microbial insecticides represents the commercial cutting edge of biotechnology.

The term insecticide usually bring thoughts of poisonous, non-selective, energy-consuming chemicals that are used to kill pest insects. However, not all insecticides fit this generally accepted stereotype. Those microorganisms which often suppress and naturally control populations or pests which injure man and causes extensive damage to food and fibre crops. Formulation of living viruses, bacteria, fungi and protozoa are called microbial insecticides.

Revolutionary techniques in biotechnology offer tantilizing new prospects for the future. Biological control has a long history, reaching into the undated past when the chinease used Pharoah's ants to control pests of stored gain. Others date the start of biological control to the domestication of the cat. The introduction of an exotic species to control a pest, the classical form of biological control, is claimed to date from 1762 when the indian Mynah was brought to mauritius to control the Red Locust. Subsequent attempts at introduction ocassionally had unwanted results, the introduction of cats to control rats on Ascension Island in 1815 resulted in the extermination of virtually all the sea birds on the island. Successful control of cotton cushion scale by Australian ladybirds in california in 1888 encouraged practitioners to continue to develop biological pest control by the introduction of exotic enemies.

Haplodiplodisation

Many species are able to produce haploids through different in vitro techniques. The oldest was anther culture of androgenesis. To date, the results vary considerably from one species to another. Solanae (datura, tobacco, red pepper, eggplant, petunia), cereals (wheat, barley, rice, triticale, maize) or crucifers (soybean, cabbage) are species easy to generate by anther culture. To the contrary, tomatoes, leguminous or compositae are recalcitrant. Ovule culture, or gynogenesis was a successful technique for Gerbera, beet, courgette. However, the most common teechnique is pollination with irradiated pollen, in order to induce in vivo parthenogenesis. The oospheres developed in embryos without fertilization are saved by embryo rescue.

Ketolactose Test For Agrobacterium Biotype 1

OBJECT: To identify specific Agrobacterium strains.

MATERIALS:

MEDIA: yeast-extract indicator medium (YI).

REAGENTS: Benedict's reagent

EQUIPMENT AND SUPPLIES: Inoculating loop, Agrobacterium biotype 1 culture, Agrobacterium biotype 2 culture, Escherichia coli culture.


PROCEDURE:

1. With the help of waterproof marker or wax pencil, draw lines on the bottom of a YI plate to divide it into three pie-shaped sections of equal size.

2. Use flame-sterilized inoculation loop, streak one third of the plate with Agrobacterium biotype 1 cells, one third with biotype 2 cells and the last third with E.coli cells.

3. Mention your name, the type of cell in each section of the plate and the date on the bottom of the plate.

4. Incubate the plate in an inverted position for two days at 28-29 'c.

5. Flood the YI plate with 4 ml of Benedict's reagent.

6. Incubate the plate at room temperature for one hour.

Following is a simple technique for isolating and precipitating high molecular nuclear DNA from plant cells. A mortar and pestle are used to break cells open mechanically and to disrupt the plasma membrane. The plasma membrane is further degraded by the use of an extraction buffer containing a detergent that dissolves membranes. Their combined action produces a homogenate containing cell wall and plasma membrane fragments as well as intact nuclei, chloroplasts and mitochondria. Low-speed centrifugation is used to separate the nuclei from the smaller organelles.

OBJECT: To demonstrate how DNA is isolated from plant tissues.

MATERIALS: REAGENTS: Cauliflower homogenization solution, sodium citrate solution, sodium chloride
solution, Absolute ethanol.

           EQUIPMENT AND SUPPLIES: Centrifuge, Graduated cylinder, Funnel, Cheese-cloth, Centrifuge bottles, Centrifuge tubes, Chilled mortar and pestle, Glass rod, Razor blade, Fresh cauliflower head.

PROCEDURE:

1. Using a razor blade, remove 25g of the outer 2-3 mm of the cauliflower surface.

2. Place the tissue in a mortar and add 25 ml of sodium citrate solution. Grind the mixture until it becomes a smooth slurry.

3. Add 150 ml of cauliflower homogenization solution to the mortar. Continue grinding the mixture an additional 30 min.

4. Filter the homogenate through a funnel lined with two layers of cheese-cloth. Squeeze the cloth to recover any additional liquid. Transfer the liquid two centrifuge bottles.

5. Add 2 volumes of absolute ethanol to each bottle while stirring continuously. Balance the bottles and centrifuge them for 5 min at 200 x g at 4'c.

6. Pour off the supernatant, saving the pellet containing nuclei at the botton of the bottles.

7. Add 1.5 volumes of sodium chloride solution and stir the mixture.

8. Transfer the mixture to a pair of clean centrifuge tubes. Centrifuge them at 10,000 x g for 25 min at 20'c.

9. Save the supernatant in a clean beaker.Resuspend the pellet in 15 ml of sodium chloride solution.   Centrifuge the mixture at 10,000 x g for 25 min at 20'c.

10.Add the supernatant to the beaker containing supernatant from the previous centrifugation. Slowly add an equal volume of absolute ethanol while slowly stirring with a glass rod.

11. Fibrous DNA strands will collect on the rod. continue stirring until DNA no longer adheres to the rod.

Booroola Gene

Gene mapping is essential as the foundation for genetic manipulation. Thus far, however, few specific genes of significance to animal agriculture have been identified, isolated or mapped. one example of a gene that is beginning to be understood, although it has not been specifically isolated, is he BOOROOLA GENE from Australian merino sheep. This gene boosts the incidence of twinning and triplets in sheep, giving an overall 20-40 percent increase in the number of lambs weaned. Introducing the booroola gene into other sheep and cattle could offer a fast, reliable way to increase the productivity of ewe and cow herds. Although the gene could be crossed into some breeds by sexual breeding, its introduction by molecular gene transfer would be faster and more important, it would allow the trait to be passed to a wider range of livestock. Mapping of the booroola gene is helping scientists determine more precisely how the gene operates and is also aiding in its cloning. Scientists may then attempt to transfer the gene to other valuable livestock species.

DNA Chip Technology

DNA chip technology, a marriage of the semi-conductor manufacturing industry and molecular genetics, will transform genetic analysis because it allows us to analyze tens of thousands of genes simultaneously on a single chip. The manufacturing process of microchips and DNA chips is similar, in principle, but instead of shining light through a series of masks to each circuits into silicon, automated DNA chip-makers use a series of masks to lay down an array of DNA fragments on a glass slide. DNA chip technology is being used for:

1. detect mutations in disease-causing genes.

2. monitor gene activity.

3. diagnose infectious diseases and identify the best antibiotic treatment.

4. identify genes important to crop productivity.

5. improve screening for microbes used in bioremediation.

DNA chip will be essential for converting the raw genetic data provided by the Human Genome Project into useful products.

Germplasm Conservation (Gene Bank)

Tissue culture methods offer the opportunity for in vitro collecting, rapid multiplication and distribution of important elite, or rare plants that are threatened with extinction. The two major in vitro  storage strategies are slow growth and cryo-preservation. Since the first results of seibert (1976), who was able to initiate shoots from carnation shoot apices frozen to -196'c. This technique is now successful for many of horticultural species. Dereuddre et al. (1991) have provided a very simple technology to freeze encapsulated meristems in dried alginate beads. It works for pear, strawberry, eucalyptus, potato. The international Potato center (CIP) in Lima, Peru has a large word potato collection. Germplasm of sweet potato and cassava is at the International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria.

The movement of germplasm involves the risks of accidentially introducing plant quarantine pests along with the host plant material. To limit these risks, the plant material should be transferred from one country to another as in vitro cultures through a transit centre, where it should be indexed. For bananas, in the framework of INIBAP, the transit centre is the catholic university of Leuven in Belgium where a very large in vitro germplasm exists. The germplasm conservation (vitro preservation) of asexually propagated horticultural crops for storage under conditions closer to ambient temperature to suit the local conditions are important. An in vitro storage slow growth and cryopreservation have been standardized for many horticultural crops. Freezing of encapsulated meristems is possible in pears, strawberry and potato. For the movement of germplasm, in-vitro technique is very handy. The molecular taxonomy (like RAPD and RFLP) in genomic analysis and classification and development of molecular tools for disease indexing in in vitro propagated materials, in checking clonal fidelity, germplasm classification and identification of markers linked are important priority areas in this field. Molecular characterization of indigenous germplasm, application of DNA markers for identification of cultivars and molecular linkage maps for developing new varieties are important aspects of biotechnological studies.

Prime Numbers Less Than 1000

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97 101 103 017 109 113 127 131 137 139 149 151

157 163 167 173 179 181 191 193 197 199 211 223

227 229 233 239 241 251 257 263 269 271 277 281

283 293 307 311 313 317 331 337 347 349 353 359

367 373 379 383 389 397 401 409 419 421 431 433

439 443 449 457 461 463 467 479 487 491 499 503

509 521 523 541 547 557 563 569 571 577 587 593

599 601 607 613 617 619 631 641 643 647 653 659

661 673 677 683 691 701 709 719 727 733 739 743

751 757 761 769 773 787 797 809 811 821 823 827

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919 929 937 941 947 953 967 971 977 983 991 997

...and they just keep going on and on forever...

Cartagena Protocol On Biosafety

Overview of the Biosafety Protocol:

The Biosafety Protocol seeks to protect biological diversity from the potential risks posed by living modified organisms resulting from modern biotechnology.

The Biosafety Protocol makes clear that products from new technologies must be based on the precautionary principle and allow developing nations to balance public health against economic benefits. It will for example let countries ban imports of a genetically modified organism if they feel there is not enough scientific evidence the product is safe and requires exporters to label shipments containing genetically altered commodities such as corn or cotton.


Objective of the Protocol:

In accordance with the precautionary approach, contained in Principle 15 of the Rio Declaration on Environment and Development, the objective of the Protocol is to contribute to ensuring an adequate level of protection in the field of the safe transfer, handling and use of 'living modified organisms resulting from modern biotechnology' that may have adverse effects on the conservation and sustainable use of biological diversity, taking also into account risks to human health, and specifically focusing on transboundary movements.

Living modified organisms (LMOs):

Living modified organisms (known as LMOs) resulting from modern biotechnology are broadly equivalent to genetically modified organisms. 'Modern biotechnology' is defined in the Protocol to mean the application of in vitro nucleic acid techniques, or fusion of cells beyond the taxonomic family, that overcome natural physiological reproductive or recombination barriers and are not techniques used in traditional breeding and selection.


The Protocol and the Precautionary Approach:



One of the outcomes of the United Nations Conference on Environment and Development (also known as the Earth Summit) held in Rio de Janeiro, Brazil, in June 1992, was the adoption of the Rio Declaration on Environment and Development, which contains 27 principles to underpin sustainable development. Commonly known as the precautionary principle, Principle 15 states that "In order to protect the environment, the precautionary approach shall be widely applied by States according to their capabilities. Where there are threats of serious or irreversible damage, lack of full scientific certainty shall not be used as a reason for postponing cost-effective measures to prevent environmental degradation."
Elements of the precautionary approach are reflected in a number of the provisions of the Protocol, such as:

• The preamble, reaffirming "the precautionary approach contained in Principle 15 of the Rio Declaration on environment and Development";
• Article 1, indicating that the objective of the Protocol is "in accordance with the precautionary approach contained in Principle 15 of the Rio Declaration on Environment and Development";
• Article 10.6 and 11.8, which states "Lack of scientific certainty due to insufficient relevant scientific information and knowledge regarding the extent of the potential adverse effects of an LMO on biodiversity, taking into account risks to human health, shall not prevent a Party of import from taking a decision, as appropriate, with regard to the import of the LMO in question, in order to avoid or minimize such potential adverse effects."; and
• Annex III on risk assessment, which notes that "Lack of scientific knowledge or scientific consensus should not necessarily be interpreted as indicating a particular level of risk, an absence of risk, or an acceptable risk."

What does the Protocol cover?

The Protocol applies to the transboundary movement, transit, handling and use of all living modified organisms that may have adverse effects on the conservation and sustainable use of biological diversity, taking also into account risks to human health.

Parties and non-Parties to the Protocol:

The governing body of the Protocol is called the Conference of the Parties to the Convention serving as the meeting of the Parties to the Protocol (also the COP-MOP). The main function of this body is to review the implementation of the Protocol and make decisions necessary to promote its effective operation. Decisions under the Protocol can only be taken by Parties to the Protocol. Parties to the Convention that are not Parties to the Protocol may only participate as observers in the proceedings of meetings of the COP-MOP.

The Protocol addresses the obligations of Parties in relation to the transboundary movements of LMOs to and from non-Parties to the Protocol. The transboundary movements between Parties and non-Parties must be carried out in a manner that is consistent with the objective of the Protocol. Parties are required to encourage non-Parties to adhere to the Protocol and to contribute information to the Biosafety Clearing-House.

Relationship between the Protocol and the WTO:

A number of agreements under the World Trade Organization (WTO), such as the Agreement on the Application of Sanitary and Phytosanitary Measures (SPS Agreement) and the Agreement on Technical Barriers to Trade (TBT Agreement), and the Agreement on Trade-Related Aspects of Intellectual Property Rights (TRIPs), contain provisions that are relevant to the Protocol. The Protocol states in its preamble that it:

• Recognizes that trade and environment agreements should be mutually supportive;
• Emphasizes that the Protocol is not interpreted as implying a change in the rights and obligations under any existing agreements; and
• Understands that the above recital is not intended to subordinate the Protocol to other international agreements. 

Main features of the Protocol:

The Protocol promotes biosafety by establishing rules and procedures for the safe transfer, handling, and use of LMOs, with specific focus on transboundary movements of LMOs. It features a set of procedures including one for LMOs that are to be intentionally introduced into the environment called the advance informed agreement procedure, and one for LMOs that are intended to be used directly as food or feed or for processing. Parties to the Protocol must ensure that LMOs are handled, packaged and transported under conditions of safety. Furthermore, the shipment of LMOs subject to transboundary movement must be accompanied by appropriate documentation specifying, among other things, identity of LMOs and contact point for further information. These procedures and requirements are designed to provide importing Parties with the necessary information needed for making informed decisions about whether or not to accept LMO imports and for handling them in a safe manner.



The Party of import makes its decisions in accordance with scientifically sound risk assessments. The Protocol sets out principles and methodologies on how to conduct a risk assessment. In case of insufficient relevant scientific information and knowledge, the Party of import may use precaution in making their decisions on import. Parties may also take into account, consistent with their international obligations, socio-economic considerations in reaching decisions on import of LMOs.



To facilitate its implementation, the Protocol establishes a Biosafety Clearing-House for Parties to exchange information, and contains a number of important provisions, including capacity-building, a financial mechanism, compliance procedures, and requirements for public awareness and participation.




Procedures for moving LMOs across borders:

Advance Informed Agreement:

The "Advance Informed Agreement" (AIA) procedure applies to the first intentional transboundary movement of LMOs for intentional introduction into the environment of the Party of import. It includes four components: notification by the Party of export or the exporter, acknowledgment of receipt of notification by the Party of import, the decision procedure, and opportunity for review of decisions. The purpose of this procedure is to ensure that importing countries have both the opportunity and the capacity to assess risks that may be associated with the LMO before agreeing to its import. The Party of import must indicate the reasons on which its decisions are based (unless consent is unconditional). A Party of import may, at any time, in light of new scientific information, review and change a decision. A Party of export or a notifier may also request the Party of import to review its decisions.

However, the Protocol's AIA procedure does not apply to certain categories of LMOs:

• LMOs in transit;
• LMOs destined for contained use;
• LMOs intended for direct use as food or feed or for processing.

LMOs intended for food or feed, or for processing:

LMOs intended for direct use as food or feed, or processing (LMOs-FFP) represent a large category of agricultural commodities. The Protocol, instead of using the AIA procedure, establishes a more simplified procedure for the transboundary movement of LMOs-FFP. Under this procedure, A Party must inform other Parties through the Biosafety Clearing-House, within 15 days, of its decision regarding domestic use of LMOs that may be subject to transboundary movement.

Decisions by the Party of import on whether or not to accept the import of LMOs-FFP are taken under its domestic regulatory framework that is consistent with the objective of the Protocol. A developing country Party or a Party with an economy in transition may, in the absence of a domestic regulatory framework, declare through the Biosafety Clearing-House that its decisions on the first import of LMOs-FFP will be taken in accordance with risk assessment as set out in the Protocol and time frame for decision-making.

Handling, Transport, Packaging and Identification:

The Protocol provides for practical requirements that are deemed to contribute to the safe movement of LMOs. Parties are required to take measures for the safe handling, packaging and transportation of LMOs that are subject to transboundary movement. The Protocol specifies requirements on identification by setting out what information must be provided in documentation that should accompany transboundary shipments of LMOs. It also leaves room for possible future development of standards for handling, packaging, transport and identification of LMOs by the meeting of the Parties to the Protocol.

Each Party is required to take measures ensuring that LMOs subject to intentional transboundary movement are accompanied by documentation identifying the LMOs and providing contact details of persons responsible for such movement. The details of these requirements vary according to the intended use of the LMOs, and, in the case of LMOs for food, feed or for processing, they should be further addressed by the governing body of the Protocol.

Biosafety Clearing-House:

The Protocol established a Biosafety Clearing-House (BCH), in order to facilitate the exchange of scientific, technical, environmental and legal information on, and experience with, living modified organisms; and to assist Parties to implement the Protocol. It was established in a phased manner, and the first meeting of the Parties approved the transition from the pilot phase to the fully operational phase, and adopted modalities for its operations.