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Paper Information

Journal:   JOURNAL OF AGRICULTURAL SCIENCES   Fall 2004 , Volume 10 , Number 3; Page(s) 59 To 86.
 
Paper: 

ISOLATION AND CULTIVATION OF EMBRYOGENIC MICROSCOPE FROM BARLEY (HORDEUM VULGARE) FOR OPTIMAZING CULTURE CONDITIONS AND PRODUCING DOUBLED HAPLOID PLANTS

 
 
Author(s):  HASHEMI M., KHOSSROSHAHLI M., GHANADHA M.R., BOZORGIPOUR R.
 
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Abstract: 

In this study the effects of temperatures stress and manitol pretreatment on microspore cultures response and viability of microspores have been examined in three popvlations of F3 (Alpha/Yazd -5, Arigashar/Yazd -5, Arigashar/Alger) and cultivar Igri. Spikes were sampled when the distance from the base of flage leaf to the penultimate leaf was 2 to 5 cm in winter genotypes and 5 to 10 cm in spring genotypes. In cytological observation, most microspores in the anther of central floret of each spike were at the mid to late uninucleate stage. The physical appearance of the spikes at this stage depends upon the genotype and the environment where plants are grown. Three pretreatment were investigated in first study.l.cold pretreatment of spikes: Spikes were pretreated during 28 day indarkness and at 4°c. 2. pretreatment of spikes in manitol: Spikes were placed in plate with fifteen ml of 0.3 M Ice cold manitol and kept in the fridge at 4°c for 4-7 days. 3. pretreatment of anther: Fresh anthers were pretreatmented in 5 cm plates with 0.3 M manitol solution at 25°c (3-4 days) in PH=7.00. 200 anthers of central part of each spikes were constituted one replication. In this experiment mechanichal isolation blending method was used to isolate microspore. FHG liquid media (Hunter 1988) with 10 mg/lit PAA (phenyl acetic acid) was used for barley microspore culture. Genotype differences and pretreatments effect were highly significant in microspore viability and their response proportion. There was a very singnificant intraction pretreatments and genotypes according to the percentage of embryo like structure although this subject was not significant for viability of microspores. In second study the influence of temperature stress on microspore culturesresponse and their viability of microspores in cultivar Igri has been examined. Spikes were pretreated in darkness at 4°c for periods of 7,14 and 28 days. According to viability of microspores and microspore culture responses cold pretreatment differences were singnificant. In third study significant differences were observed between four mechanical microspore isolation methods: (Vortexing, Blending, Stirring and Maceration) according to the number ofreleased microspores from each anther, number of relesed viadle microsporespr anther an in number of divided microspore per anther. Finally, in forth study, the influence of concentration of PAA on microspore culture response in four genotypes has been examined. Genotype differences and various concentration of PAA were highly significant in microspores response proportion. There was a very significant intraction between genotype and hormone concentration. Attentive to results it seems that pretreatment of anthers in manitol mechanically isolation method ofvortexing, concentration of 10 mg flit PAA in FHG media are sutible in microspore culture of barley. Ina routin breeding program such as produce doubled haploid, genetic engineering, mutation and selection, the proportion of response of microspore culture is suitable enough.

 
Keyword(s): DOUBLED HAPLOID, MICROSPORE CULTURE, EMBRYOGENIC MICROSCOPE, HORDEUM VULGARE, PAA (PHENYL ACETIC ACID)
 
References: 
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