大麦单粒种子起源的小孢子培养再生频率研究

Study on the Regeneration Frequency of Isolated Microspores Derived from a Single-Seed of Barley

为提高单株小孢子培养的再生效率,以大麦品种花30为材料,在人工气候室研究了离体穗、离体小花预处理时间和诱导培养基中无机氮、有机氮含量以及添加PEG对愈伤组织产量、绿苗产量和再生能力的影响。结果表明,离体穗5℃处理19 d的愈伤组织和绿苗产量及再生能力均优于32 d的,离体小花预处理1～2 d可促进愈伤组织的形成和绿苗的分化;降低无机氮浓度[KNO_3降至707.5 mg·L^(-1),(NH_4)_2SO_4降至115.75 mg·L^(-1)]可提高愈伤组织和绿苗产量及再生能力;提高有机氮浓度(谷氨酰胺从400 mg·L^(-1)提高到1 600 mg·L^(-1),水解酪蛋白从100 mg·L^(-1)提高到400 mg·L^(-1))可增加小孢子愈伤组织和绿苗产量;培养基中添加4%PEG可提高小孢子愈伤组织产量和质量。

In order to improve the regeneration frequency of isolated microspores derived from a single-seed of barley,a single donor plant grown in chamber for barley cultivar Hua 30,and the effects of the period of pretreatment on young spikes and floret,the content of inorganic and organic nitrogen in induction medium,and the addition of PEG on the formation of callus and green plants were studied. The results showed that the pretreatment of 19 d at 5℃ on young spikes worked better than the pretreatment of 32 d. The pretreatment on floret for 1-2 d benefited the formation of callus and green plants. Low level of inorganic nitrogen in induction medium[KNO 3 reduced to 707.5 mg·L -1 and(NH 4) 2SO 4 reduced to 115.75 mg·L -1 ] improved the production of both callus and green plants. The elevation of organic nitrogen level(casein glutamine increased from 400 mg·L -1 to 1 600 mg·L -1 ,and hydrolysate increased from 100 mg·L -1 to 400 mg·L -1 ) promoted the callus formation and green plant regeneration. In addition,the supplement of 4% PEG also improved the quantity and quality of callus formation.