Research Center of Agricultural Biotechnology, Ningxia Academy of Agricultural and Forestry Sciences, Yinchuan, Ningxia, China.
College of Agriculture, Gansu Agricultural University, Lanzhou, Gansu, China.
PeerJ. 2023 May 9;11:e14984. doi: 10.7717/peerj.14984. eCollection 2023.
As the primary means of plant-induced haploid, anther culture is of great significance in quickly obtaining pure lines and significantly shortening the potato breeding cycle. Nevertheless, the methods of anther culture of tetraploid potato were still not well established.
In this study, 16 potato cultivars (lines) were used for anther culture . The corresponding relation between the different development stages of microspores and the external morphology of buds was investigated. A highly-efficient anther culture system of tetraploid potatoes was established.
It was shown in the results that the combined use of 0.5 mg/L 1-Naphthylacetic acid (NAA), 1.0 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D), and 1.0 mg/L Kinetin (KT) was the ideal choice of hormone pairing for anther callus. Ten of the 16 potato cultivars examined could be induced callus with their respective anthers, and the induction rate ranged from 4.44% to 22.67% using this hormone combination. According to the outcome from the orthogonal design experiments of four kinds of appendages, we found that the medium with sucrose (40 g/L), AgNO (30 mg/L), activated carbon (3 g/L), potato extract (200 g/L) had a promotive induction effect on the anther callus. In contrast, adding 1 mg/L Zeatin (ZT) effectively facilitated callus differentiation.
Finally, 201 anther culture plantlets were differentiated from 10 potato cultivars. Among these, Qingshu 168 and Ningshu 15 had higher efficiency than anther culture. After identification by flow cytometry and fluorescence hybridization, 10 haploid plantlets (5%), 177 tetraploids (88%), and 14 octoploids (7%) were obtained. Some premium anther-cultured plantlets were further selected by morphological and agronomic comparison. Our findings provide important guidance for potato ploidy breeding.
作为植物诱导单倍体的主要手段，花药培养对于快速获得纯系和显著缩短马铃薯育种周期具有重要意义。然而，四倍体马铃薯花药培养的方法仍未得到很好的建立。
本研究以 16 个马铃薯品种（系）为材料进行花药培养，研究了小孢子不同发育阶段与花蕾外部形态的对应关系，建立了高效的四倍体马铃薯花药培养体系。
结果表明，0.5mg/L1-萘乙酸（NAA）、1.0mg/L2,4-二氯苯氧乙酸（2,4-D）和 1.0mg/L 激动素（KT）组合使用是花药愈伤组织诱导的理想激素组合。在 16 个供试马铃薯品种（系）中，有 10 个品种（系）的花药可以诱导愈伤组织，激素组合诱导率为 4.44%~22.67%。通过 4 种附加物的正交设计实验，发现培养基中添加 40g/L 蔗糖、30mg/L 硝酸银、3g/L 活性炭、200g/L 马铃薯提取物对花药愈伤组织有促进诱导作用，而添加 1mg/L 玉米素（ZT）则能有效促进愈伤组织分化。
最终从 10 个马铃薯品种（系）中分化出 201 个花药培养植株，其中清水 168 号和宁薯 15 号的花药培养效率高于对照。经流式细胞仪和荧光杂交鉴定，获得 10 个单倍体植株（5%）、177 个四倍体植株（88%）和 14 个八倍体植株（7%）。通过形态学和农艺性状比较，进一步筛选出一些优质花药培养植株。本研究结果为马铃薯倍性育种提供了重要指导。
