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)
為了明確鹽預處理對大麥種子萌發和小孢子培養的影響，本研究以耐鹽品種‘花11’和鹽敏感品種‘花30’為材料，比較了它們在不同鹽濃度預處理下，種子萌發過程中的發芽率、主根長以及小孢子培養過程中的愈傷組織產量差異，同時對2個品種在小孢子時期響應鹽脅迫的4個關鍵基因的表達進行了研究。結果表明：‘花30’的種子和小孢子在無NaCl和低濃度NaCl預處理時，對鹽脅迫的耐受性均低于‘花11’，但經高濃度NaCl (萌發期為15 g/L，小孢子時期為300 mg/L)預處理后，‘花30’的種子和小孢子對高濃的耐受性均顯著增強，且發芽率、主根長和愈傷產量均顯著高于‘花11’；通過對2個品種在鹽預處理下的發芽率、主根長以及愈傷產量相對值的相關性分析發現，2個品種在種子萌發期和小孢子時期響應鹽預處理的相對值呈顯著正相關。同時，在小孢子培養階段，經過300 mg/L NaCl預處理后，‘花30’中小熱激蛋白基因HvsHSP1和翻譯起始因子1基因HveIF1A的表達量在高濃度鹽脅迫下較‘花11’顯著上調。因此，供試品種間在種子萌發過程和小孢子培養過程對鹽預處理的響應差異在某種程度上可能具有一致性，基因HvsHSP1和HveIF1A可能與鹽敏感品種在小孢子時期響應NaCl鹽誘導存在關系。
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.