| Anther culture is one of the efficient ways to establish regeneration system in haploid culture,and it could simplify the breeding process,speed up the haploid breeding advancement when combining with the traditional breeding way.The research about the tissue culture of the balsam pear(Momordica charantia L.) was not so much,and mainly concentrated on the stem tip culture and the cotyledon induction.In this research,the anther induction culture of three kinds of balsam pears(Momordica charantia L.) was studied,and it was refering to the effect of the genotype,the different auxin and its content, microspore developmental stages,the time of cold pretreatment,and different carbon and its concentration on induction of anthers.Besides,the calluses of anthers were departed to different type and their regeneration was studied.The main results were as follows:1.The relativity between buds' size and the microspores' developmental stages:The 2.5～3.5mm long buds were in tetrad stage,the 3.5～4.5mm long buds were in early-uninucleate stage,the 4.5～6.0mm long buds were in late-uninucleate stage.2.The result showed that each variety had the highest inductivity of callus in MS medium with 2,4-D 2.0 mg/L and 6-BA 2.0 mg/L,as the Bixiu's 65.91%,Dabai's 82.14%and Changbai's 94.74%.6-BA 2.0 mg/L was better for Bixiu and Dabai to form callus than 6-BA＞2.0 mg/L or 6-BA＜2.0 mg/L.Changbai could keep a high callus formation rate even when 6-BA 3.0mg/L,2,4-D 1.0mg/L and 2.0mg/L.The change of 2,4-D's content could obviously effect the callus formation rate.As the content of 2,4-D increased,the callus form faster and more,but the callus get brown faster too.3.The callus formation rate of late-uninucleate was notable higher than other microspore developmental stages.In this stage,Bixiu's callus formation rate was 79.74%,Dabai's callus formation rate was 74.94%,and Changbai's callus formation rate was 89.38%. 4.Oppose to many other research results of anther culture,in this research,pretreatment under 4℃appeared bad effect.The fresh anthers formed more calluses than the anthers had been pretreatment under 4℃.The callus formation rate decreased as the pretreatment time increased,especially when the time was 4days,the rate of Bixiu callus formation from 53.19%reduced to 18.18%,Dabai's from74.82%reduced to 24.38%,Changbai's from 88.40%reduced to 26.83%.5.The sucrose was better for the anther culture than the glucose.Bixiu's callus formation rate get the highest 83.33%when the concentration of sucrose was 3%,and as the sucrose increased,the rate go down gradually.However,as to Dabai and Changbai,the rate get to highest 70.23%and 72.73%when the sucrose was 5%.When use the glucose as the carbon, each variety form much less callus.Especially when the glucose was 7%,the rate of Changbai and Dabai both below 10%and Bixiu even could not form callus.6.The callus could be separated to two types:one type was yellow white colour with strumae,which could turn green quickly and grown fast in the inducing culture medium, and it grown well especially in treatment(2) with 6-BA2.0mg/L,KT1.0 mg/L, NAA0.2mg/L.The other type was yellow white colour too,but without strumae,looked brilliant,which could not turn green easily and grown slowly,and it get a little green in treatment(1) with 6-BA3.0 mg/L,KT1.0 mg/L,maltose 30g/L.And in treatment(1) with 6-BAI.0mg/L,KT1.0 mg/L,NAA1.0 mg/L,it could keep its texture well,although not green.7.Proper kind of auxin and its content was key to the development of callus.Increasing the suitable content of 6-BA could be effective for the callus to turn green and grow.NAA was important for the conservation of yellow green callus.ZT didn't work effectively on the induction of callus.Some of Changbai's callus formed roots in treatment(1) with 6-BA1.0mg/L,KT1.0 mg/L,NAA1.0mg/L. |
