第一例栽培种花生花药培养单倍体植株的创制

The first report on one haploid plant in cultivated peanut using anther culture technique

以栽培种花生为材料,开展花生单倍体培养的前沿性探索,包括离体花药诱导形成愈伤组织、愈伤组织分化形成幼芽、再生苗培养、再生植株倍性鉴定和嫁接移植等研究。通过比较外植体灭菌时间、诱导培养基、分化培养基、再生培养基和再生植株创制培养条件等试验,筛选出适合花生花药培养的方法:1%NaClO消毒灭菌9 min,愈伤组织的诱导培养采用B5N1、B3N1培养基,再分化培养采用B5N1-2培养基,再生苗培养采用SG培养基。本研究获得1株花药培养的再生植株,编号为15B8-8。荧光原位杂交技术(FISH)鉴定结果表明,该植株具有20条染色体,其中9条为A染色体、11条为B染色体,是第一例来自栽培种花生花药培养的单倍体植株;研究还表明,汕油52花生品种具有较强的花药培养力,有潜力作为花生花药培养的“桥梁品种”。

Cultivated peanut was used to develop haploid plant including dedifferentiation of detached anthers for callus induction,callus differentiation to form shoot,induction of adventitious shoots and roots,identification of plant ploidy and transplantation.By comparison of sterilization period,different additives and media for cali induction and shoot induction,and plant regeneration,a protocol suitable for peanut anther culture was established including 1%NaClO disinfection for 9 min,dedifferentiation induction culture using B5N1 or B3N1 medium,differentiation culture using B5N1-2 medium,plant regeneration using CM medium or SG medium.One plant,named 15B8-8,were first obtained from cultivated peanut through anther culture.Result from fluorescence in situ hybridization(FISH)showed that 15B8-8 was the target haploid line for having 20 chromosomes with 9 A chromosomes and 11 B.The result indicated that Shanyou 52 had stronger ability of anther culture,which can be considered as a bridge variety for anther culture of peanut in the future.