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逆境下植物大量积累脯氨酸是减轻胁迫伤害的一种自我保护机制。本研究应用同源克隆方法从NaCl处理的野生大豆中克隆获得一个脯氨酸合成酶(P5CS)基因,命名为GsP5CS。该基因核苷酸序列全长2.232 kb,含一个2148bp开放阅读框,编码715个氨基酸,包含有高等植物P5CS蛋白质的5个主要功能域,与菜豆PvP5CS1基因核苷酸序列相似性高达98.79%。Real Time PCR分析显示该基因受轻度盐胁迫诱导上调表达,根中表达高峰出现在200 mmol/L NaCl处理下,相对表达量为对照的5.83倍;叶片中表达高峰出现在300 mmol/L NaCl处理条件下,相对表达量为对照的12.78倍。并且该基因在根和叶片中的表达模式和脯氨酸含量的变化模式相同。上述结果说明,GsP5CS可能参与野生大豆脯氨酸合成。
Proliferation of plants under heavy stress is a self-protection mechanism to reduce stress damage. In this study, homologous cloning method was used to clone a P5CS gene from NaCl-treated wild soybean and named it as GsP5CS. The nucleotide sequence of this gene was 2.232 kb in length and contained a 2148 bp open reading frame encoding 715 amino acids. It contained five major functional domains of the P5CS protein of higher plant, which shared 98.79% similarity with the nucleotide sequence of PvP5CS1. Real Time PCR analysis showed that the gene was up-regulated by mild salt stress, and the highest peak appeared in the roots at 200 mmol / L NaCl treatment, the relative expression level was 5.83 times of that of the control; the peak expression in the leaves appeared in 300 mmol / L NaCl Under the treatment conditions, the relative expression of 12.78 times the control. And the expression pattern of this gene in roots and leaves is the same as that of proline content. The above results indicate that GsP5CS may be involved in the synthesis of wild soybean proline.