茄子小孢子培养技术研究
Microspore culture technology for eggplant
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摘要: 目的开展茄子小孢子培养技术研究,为茄子单倍体育种在生产实践上应用提供技术支持.方法以6个不同基因型茄子栽培品种为试材进行小孢子培养,探究基因型、花药预处理、花药壁组织及供体植株生长环境对茄子小孢子培养的影响.结果6个茄子栽培品种均能诱导出愈伤组织,但诱导频率差异明显,其中,37号和128号品种的小孢子愈伤组织诱导频率极显著高于其他品种(P< 0.01,下同);6个茄子品种中仅37号品种能形成再生植株,其中5株为单倍体植株,8株为双单倍体植株.茄子小孢子培养必需进行高温(36℃)热激处理,最佳处理时间6d;低温(4℃)预处理不能启动茄子小孢子脱分化形成愈伤组织,但能极大促进愈伤组织形成;花药预处理以低温(4℃)处理6d+高温(36℃)处理6d的效果最佳,愈伤组织的诱导频率达60.0个/花药;花药壁组织与小孢子共培养能促进愈伤组织形成,最佳处理时间8~12 d.供体植株在露地环境条件下小孢子愈伤组织诱导频率极显著高于温室环境条件.结论对露地环境条件下种植的37号和128号茄子的花药进行低温(4℃)处理6d+高温(36℃)处理6d,可获得最佳的小孢子愈伤组织诱导效果.Abstract: Objective In this study,the technology of microspore culture of eggplant was studied to provide technical support for the application of haploid breeding of eggplant,Method Six eggplant varieties with various genotypes were used as experimental materials to conduct microspore culture.The effects of factors such as genotype,anther pretreatment,anther wall tissue and donor plant development environment on microspore culture were studied.Result Callus could be induced from the six cultivated varieties.But callus induction frequency was very different.Callus induction frequencies of varieties No.37 and No.128 were extremely significantly higher than those of other varieties (P<0.01,the same as below).Regenerated plants were obtained from variety No.37 only,with five haploid plants and eight double haploid plants.High temperature (36 ℃) heat shock treatment was essential to microspore culture of eggplant,the best treatment time was 6 d.Low temperature pretreatment (4 ℃) could not induct callus through dedifferentiation of microspores,but it could greatly improve callus induction frequency.Anther pretreatment with low temperature(4 ℃) for 6 d plus high temperature (36 ℃) treatment for 6 d had the optimal effects,and callus induction frequency was 60.0 callus/anther.Co-cultivation of microspores with anther wall tissue improved callus induction frequency,and the best treatment time was 8-12 d.Callus induction frequency of donor plants in field condition was extremely significantly higher than that in greenhouse conditions.Conclusion Low temperature (4 ℃) for 6 d+high temperature (36 ℃) for 6 d in field condition on anther of eggplant varieties No.37 and No.128 can achieve the best microspore callus induction effect.