Shanghai Agriculture Applied Technology Development Program（2019-02-08-00-08-F01109）; Earmarked fund for China Agriculture Research System (CARS-05-01A-02); Jump-Start Program of Shanghai Academy of Agricultural Sciences （ZP19211）; Shanghai Academy of Agricultural Sciences: Program for prominent teams (Grant No. 2017B01)
为了明确盐预处理对大麦种子萌发和小孢子培养的影响，本研究以耐盐品种‘花11’和盐敏感品种‘花30’为材料，比较了它们在不同盐浓度预处理下，种子萌发过程中的发芽率、主根长以及小孢子培养过程中的愈伤组织产量差异，同时对2个品种在小孢子时期响应盐胁迫的4个关键基因的表达进行了研究。结果表明：‘花30’的种子和小孢子在无NaCl和低浓度NaCl预处理时，对盐胁迫的耐受性均低于‘花11’，但经高浓度NaCl (萌发期为15 g/L，小孢子时期为300 mg/L)预处理后，‘花30’的种子和小孢子对高浓的耐受性均显著增强，且发芽率、主根长和愈伤产量均显著高于‘花11’；通过对2个品种在盐预处理下的发芽率、主根长以及愈伤产量相对值的相关性分析发现，2个品种在种子萌发期和小孢子时期响应盐预处理的相对值呈显著正相关。同时，在小孢子培养阶段，经过300 mg/L NaCl预处理后，‘花30’中小热激蛋白基因HvsHSP1和翻译起始因子1基因HveIF1A的表达量在高浓度盐胁迫下较‘花11’显著上调。因此，供试品种间在种子萌发过程和小孢子培养过程对盐预处理的响应差异在某种程度上可能具有一致性，基因HvsHSP1和HveIF1A可能与盐敏感品种在小孢子时期响应NaCl盐诱导存在关系。
In order to definite the effect of salt pretreatment on barley seed germination and microspore culture, the salt tolerant cultivar ‘H11’ and salt sensitive cultivar ‘H30’ were used as materials and the germination rate, taproot length during the seed germination as well as the callus yield in the microspore culture stage were compared under the different salt pretreatment conditions. Meanwhile, the expression of four key genes to salt stress at the microspore stage were also investigated in the two cultivars. The results showed that the seeds and microspores of ‘H30’ were less tolerant to salt than ‘H11’ under 0 g/L NaCl and low NaCl pretreatment conditions, but under the high concentration of NaCl pretreatment (15 g/L NaCl in germination stage and 300 mg/L in microspore stage), the salt tolerance of ‘H30’ was significantly enhanced, and the germination rate, taproot length as well as callus yield with high concentration of salt stress were significantly superior to that of ‘H11’. Through the correlation analysis of germination rate, taproot length and callus yield between the two cultivars, it was found that the relative values of the differences between the two cultivars at the germination stage and the microspore stage were significantly positively correlated. Moreover, in the microspore culture stage, after 300 mg/L NaCl pretreatment, the genes HvsHSP1 and HveIF1A of ‘H30’ were significantly up-regulated in comparison with ‘H11’ under higher concentration of salt stress. It is concluded that the differences in the response of tested barley cultivars to salt pretreatment during seed germination and microspore stage may be consistent. HvsHSP1and HveIF1A of salt sensitive cultivar may be related with the response of NaCl induction in the microspore stage.