马尾松胚乳愈伤组织发生的初步研究

为获取马尾松(Pinus massoniana)单倍体材料,建立其胚乳组织培养体系,对马尾松胚乳不同的消毒方法进行筛选,并分析不同基本培养基、生长调节剂配比和热激处理对愈伤诱导的影响。结果表明,胚乳的最佳消毒方式为75%酒精处理30 s+2%NaClO消毒15 min,外植体污染率为0,死亡率仅为15.56%;生长素可促进愈伤诱导且是必需的;细胞分裂素浸泡种子可代替培养基中的细胞分裂素,并能取得比较好的胚乳愈伤诱导效果,1.0 mg·mL^(-1) 6-BA浸泡30 min处理的效果最佳,在培养基WPM+NAA 2.0 mg·L^(-1)+2,4-D 1.0 mg·L^(-1)+蔗糖30 g·L^(-1)+琼脂7 g·L^(-1)中,愈伤诱导率高达87.78%;培养基DCR+NAA 2.0 mg·L^(-1)+6-BA 1.0 mg·L^(-1)+蔗糖30 g·L^(-1)+琼脂7 g·L^(-1)的胚乳愈伤诱导率最高(68.75%);热激处理可提高胚乳愈伤的诱导率和质量,以相对湿度85%下45℃热激10 min最佳,总愈伤诱导率为66.25%,成团愈伤诱导率达8.75%;增殖培养以培养基WPM+NAA 0.1 mg·L^(-1)+6-BA 2.0 mg·L^(-1)+蔗糖30 g·L^(-1)+琼脂7 g·L^(-1)最佳,增殖率达83.33%。

A Preliminary Study on Occurrence of Endosperm Callus in Pinus massoniana

In order to obtain haploid materials of Pinus massoniana and establish endosperm tissue culture system, different disinfectants and disinfection times of P. massoniana were screened, and the effects of different hormone ratios and different heat shock temperatures and times on callus induction were studied. The results showed that the best way to sterilize explants was 75% alcohol treatment for 30 s+2% NaClO for 15 min, the explant contamination rate was 0, the mortality rate was only 15.56%; Auxin could promote callus induction and was necessary; The seeds soaked with cytokinin could replace the cytokinin in the medium, and could obtain better effect of endosperm callus induction, 1.0 mg·mL-1 6-BA for 30 min was the best, in the medium WPM+NAA 2.0 mg·L-1+2,4-D 1.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1 the induction rate was as high as 87.78%; The medium DCR+NAA 2.0 mg·L-1+6-BA 1.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1 had the highest endosperm callus induction rate (68.75%); Heat shock treatment could improve the induction rate and quality of endosperm callus. The best heat shock at 45 ℃ under 85% relative humidity for 10 minutes, the total callus induction rate was 66.25%, and the mass callus induction reached 8.75%; The best proliferation culture was WPM+NAA 0.1 mg·L-1+6-BA 2.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1, and the proliferation rate was 83.33%.