m Low temperatures are among the most important

environmental factors influencing plant distribution,

development and survival.

m Many plants from temperate regions have developed

natural adaptive mechanisms to tolerate low and
freezing temperatures.

m p


plants can adjust their metabolism
to cold and increase their tolerance to freezing in
response to low-non-freezing temperatures (Levitt,
1980).
p  ²



m „ccurs at low ÷akes place at high extent

temperatures.
Ice deposition in
m Results from loss of intracellular and
function of extracellular spaces.
biomembranes
connected with decrease
in their fluidity and
inactivation of the
membrane-bound ion
pumps
m ²rost hardening is a quality of a individual plant and is governed by
its genetic potential as well as by environmental factors, thus,
usually changes with time.

m It is difficult to be achieved by classical breeding or gene transfer

methods because 
  
conferred by the
product of one gene, but is a syndrome.

m It comprises quite different traits of cell biology, such as à
 à

    
  à



 

   
  à  
    

m Low temperature,
m Length of photoperiod.

m ÷he effect of these factors have been checked for plants

such as S
  


   

  

² 
 


m Æith gradual increase in temperature, plants deharden at

specific rates.

m It is less dependant on photoperiod than hardening.

m Inhibited photosynthesis, slower carbohydrate
translocation,

m lower respiration rates and increased protein degradation

rates.

m Impairement of integral membrane proteins involved in

transport of K+ ions .

m Intracellular ice formation is inevitably lethal, causes efflux

of Ca+2 ions.

m Ca+2 ions play important role for recovery of freeze

stressed cells.
m 2 types of survival strategies:

²  p
through:

‡ Desiccation of plant tissues
‡ Supercooling

² 
p
through:
‡ ÷olerance of freeze-induced cellular dehydration via
accumulation of various solutes in cell sap

g ~oth processes operate simultaneously in plants such as

induced in 
   
g ~ut it also depends on species-specific properties and
growth conditions.
m Éxhibited by rice and wheat seeds, dried tubers
which can sustain loss of tissue water without
loosing viability.

m Remaining water is so tightly bound that it can not

crystallize even if cooled down to -196 degree C.
u
  
m Éxhibited in temp. range -0.6 to -15 0c when extracellular ice
crystal formation takes place.
m Varies with tissue moisture content.

S 
÷emp. range -16 to -25 0c
÷ightly packed cells and reduced intraacellular spaces allow
supercooling in cold acclimated plants

 
²avoured by loss of water during cold acclimation by small cells.
Specifically parenchymal cells survive by tolerance to freeze-
induced dehydration process.
Solutes such as soluble sugars (sucrose, glucose, raffinose, fructose,
etc.) in the vacuoles improve the water holding capacity of cells and
enables avoidance of stress caused by freeze-induced dehydration.

Presence of solutes increase the osmotic potential of cytoplasm and in

turn, decrease the freezing point.

÷hus enables plant material to persist in super cooled state even at

low subzero temp.
m ¯dequate amount of cryoprotective solutes enable
tolerance of freeze-induced dehydration stress.

m Sucrose is one o the most essential cryoprotectant

and osmolyte.

m „thers include low molecular weight sugars, sugar

alcohols, free ¯mino acids, organic acids and
soluble proteins.
V
²²


S 
!²S"

m ÷hey have the ability to adsorb onto the surface of ice crystals
and inhibit their growth and recrystallization.

m It is also observed that hormone Éthylene is involved in

regulating antifreeze activity in respond to cold.

m ¯²Ps are homologous to pathogenesis-related proteins and also

provide protection against psychrophilic pathogens.

m It has been observed that transferring genes which encode

for¯²Ps, to freezing-sensitive plants will lower their freezing
point.
m ¯²Ps have been screened in plants 





m Some cold-inducible genes have been shown to
be involved in freezing tolerance .

m ¯lthough the expression of these genes seems to

be specifically regulated by low temperatures, in
most cases it is also responsive to dehydration,
salt stressand abscisic acid (¯~¯) (÷homashow,
1999), indicating that plant responses to all these
treatments share common features.
m Some cold-inducible genes such as
#$%p %  p   from
¯rabidopsis have been isolated and screened.

m Most of these genes are not uniformly regulated

by cold, suggesting that the molecular
mechanisms of cold acclimation are differentially
modulated in different tissues.
m ÷wo cold-specific-inducible genes, p!à  p

 
 "  p
, encoding multifunctional
regulatory proteins were isolated (¯barca et al., 1999;
Jarillo et al., 1994).

m In addition p#  p#
, two genes whose
expression is transiently induced by cold and in response
to drought, salt stress and ¯~¯, were also identified (Capel
et al., 1997; Medina et al., 2001).

m 
  

&  à 

p
 
  p'

à (
m Isolated by screening a cDN¯ library prepared from cold-
acclimated seedlings of ¯rabidopsis.

m Its expression is restricted to specific root cell types.

m Éncodes an active cationic peroxidase.

m  à p'  


  à   


  à p' 
 
à    (

m 
    




(
m ¯nother gene, S p  à 
 

 
 is involved in proline

synthesis, and the 
   à
 
 àdrought as well as frost resistance.
)*V(