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为了建立单株起源的小孢子高频再生体系用于突变体库创制和筛选研究,以人工气候室种植的大麦为供体材料,研究了不同培养批次和不同单株起源对小孢子愈伤诱导和植株再生频率的影响,以及盐胁迫(500 mg·L~(-1) NaCl)对单株起源小孢子培养阶段的筛选效果。结果表明,大麦单株来源的小孢子的愈伤诱导产量在不同培养批次中出现较大的波动性,抽穗进入第15周取材其愈伤产量获得最高,通过连续培养单株小孢子平均成苗2 035株;盐胁迫对单株小孢子的愈伤诱导和植株再生存在显著的抑制作用,单株平均愈伤产量和绿苗再生率与对照相比分别下降了46%和67%,不同单株之间的小孢子再生植株频率存在较大差异。结论:利用人工可控条件种植的大麦单株,通过连续小孢子培养可以获得高频率纯合再生植株;盐胁迫可以在小孢子培养阶段实现有效筛选。
In order to establish a single-source high-frequency microspore regeneration system for mutant library creation and screening studies, using barley planted in the artificial climate chamber as the donor material, we studied the effects of different culture batches and the origin of different microspores on microspore callus Induction and regeneration frequency of plantlets, and the effects of salt stress (500 mg · L -1 NaCl) on the microspore culture stage. The results showed that the callus induction yield of microspore derived from single plant of barley showed greater volatility in different culture batches. The highest callus yield was obtained in the 15th week after heading. The average number of microspore per plant There were 2 035 seedlings in each group. Salt stress significantly inhibited callus induction and plant regeneration of single plantlets. The average callus per plant and the regeneration rate of green plantlets decreased by 46% and 67%, respectively, compared with the control There was a big difference in the frequency of microspore regeneration plants between plants. CONCLUSION: High frequency homozygous regenerated plants can be obtained through continuous microspore culture using barley cultivated in artificially controllable conditions. Salt stress can be effectively screened during microspore culture.