甘蓝型油菜NER游离小孢子培养优化体系的建立(摘要)(英文)

目的]建立甘蓝型油菜NER游离小孢子培养优化体系。方法]对甘蓝型油菜新胞质不育恢复系(NER)进行游离小孢子培养,并对小孢子培养的影响因素作了初步研究。将所取材料接种在NLN-13诱导培养基上培养,并统计总胚产量和子叶胚产量,比较不同基因型间胚状体诱导效果将诱导培养基NLN-13中添加0.05mg/L6-BA、0.05mg/LNAA及二者混用,分别统计平均总胚产量和平均子叶胚产量。研究液体单层悬浮培养和固液双层培养2种培养方式对培养方式对小孢子胚产量的影响;在诱导培养基中加入不同浓度的秋水仙碱,对平均胚产量作调查。结果]不同遗传背景下的甘蓝型油菜NEA不育胞质恢复系(NER)胚产量差异显著,说明在相同试验条件下基因型的差异是造成胚产量差异的重要原因。提高小孢子胚发生能力试验表明,在诱导培养基上添加适当的6-BA和NAA有利于小孢子胚的发生;固液双层培养优于液体单层培养;在诱导培养基中添加秋水仙碱加倍处理小孢子胚产量与对照差异不明显。在诱导直接成苗前对子叶型胚进行12h/12h弱光照1～2d、在生根培养基中添加一定浓度的NAA等处理能提高小孢子胚成苗率。结论]建立了一套新胞质不育材料的小孢子培养优化体系,为加速新型胞质不育恢复系材料NER转育后代基因型纯合奠定了基础。

Establishment on Isolated Microspore Culture Optimized System for Restorer of New Cytoplasmic Male Sterile (NER) of Brassica napus L.

Objective]The aim was to establish optimized system for NER isolated microspore culture of Brassica napus L.. Method]Twenty varieties of NER of Brassica napus were grown under uncontrolled temperature and light conditions,and their isolated microspore from anthers were used as explants in vitro culture. The influencing factors of microspore culture were preliminarily studied. Result]The difference of genotypes was important influencing factors to embryoid yield. The embryoid yield increased by supplementing with 6-BA and NAA,culturing in solid-liquid double layer medium with activated charcoal; The difference was not significant of embryoid yield between culturing in medium supplemented with colchicines and the CK. The rates of cotyledonous embryoids directly developed into normal plantlets increased through enriching with 0.1-0.2 mg/L NAA and being treated on slim illumination two days before being inducted into normal plantlets. Conclusion]The technical system of microspore culture of restorer of new cytoplasmic male sterile （NER） was established,which and lays a foundation for accelerating genotype purification of NER introgressive line.