Study On Culture Of Anther And Isolated Microspore In Peanut(arachis Hypogaea L.)

New mentality and method were offered by the progress of biotechnology for peanut breeding. The breeding progress was accelerated because of apply of anther culture and isolated microspore culture technology, and it had made the foundation of haploid breeding and genetic reseach. However, anther and microspore culture of peanut has been rarely reported. Researchers studied anther culture of wild peanut, however, the callus were obtained and the hapliod plant hadn’t been acquired. 22 peanut cultivars were used as meterials in this paper. Anther and isolated microspore were incubated. Meanwhile, the growth stages of microspore, vaccination, incubation, subculture, differentiation of callus were all studied. In order to estabish the efficient induction anther culture system, theoretical basis for DH line and hapliod breeding.The results were as follows:1 Relationship of microspore development periods and induction rate(1) The isolated microspore was at tetrad stage when the length of flower bud diameter between 1.5～3.0mm; the development stage was mainly mononuclear stage while 3.1~4.7mm, and the development stage was mainly dikaryon stage while 4.8~10mm.(2) Growth result showed the induction rate of callus in tetrad, late-uninucleate and binucleate were respectively 0.83%, 30.42% and 14.58%; while microspore culture, the induction rate was 9.38%.2 Screen of anther culture condition(1) Different genotypic materials had different callus induction rate. Among 22 materials, callus of Xinghua 4 was induced easily, and induction rate was 30.42%.(2) Suorose concentration was 3%. In improved MS medium of 1/2 MS+0.8 mg/L NAA+8 mg/L 6-BA+0.02 mg/L KT, the induction rate had the highest inductivity, which was 26.25%.(3) Induction effect on hormone combinations and kinds from callus, and among the combinations of 6-BA,NAA,IBA,TDZ,KT, induction effect preferably in MS medium added 8.0mg/L 6-BA,0.8mg/L NAA,0.02mg/L KT,induction rate was high. Callus induced green was strong, and compact. In MSB5 medium added 1.0 mg/L 6-BA and 0.5 mg/L NAA, stems and roots were differentiated. Differention rate was 3%, which was higher differentiation rate than other hormone combinations.3 Screen of isolated microspore culture condition(1) Variety of Silihong was induced easiest among the all materials, and late-uninucleate was best period for materials tested,whose induction rate could reach 9.38%; embryo produced by variety of Silihong was maximum,was 0.1 per bud.(2) Induction rate of isolated microspore foster was 9.38% in MS and 1.0% in NLN fluid nutrient medium.(3) In MS fluid nutrient medium added 4 mg/L 2,4-D, 3 mg/L 6-BA and 0.3 mg/L NAA, the induction rate of microspore reached 16.7%.(4) Measuring microspore density 2.5×10~4/mL with hemocyte counting board,callus induction rate of Silihong was 12.5%.