Genomics and Biotechnology, Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada London, ON, Canada.
Front Plant Sci. 2013 Mar 25;4:60. doi: 10.3389/fpls.2013.00060. eCollection 2013.
Common bean (Phaseolus vulgaris) and black gram (Vigna mungo) accumulate γ-Glutamyl-S-methylcysteine and γ-Glutamyl-methionine in seed, respectively. Transcripts were profiled by 454 pyrosequencing data at a similar developmental stage coinciding with the beginning of the accumulation of these metabolites. Expressed sequence tags were assembled into Unigenes, which were assigned to specific genes in the early release chromosomal assembly of the P. vulgaris genome. Genes involved in multiple sulfur metabolic processes were expressed in both species. Expression of Sultr3 members was predominant in P. vulgaris, whereas expression of Sultr5 members predominated in V. mungo. Expression of the cytosolic SERAT1;1 and -1;2 was approximately fourfold higher in P. vulgaris while expression of the plastidic SERAT2;1 was twofold higher in V. mungo. Among BSAS family members, BSAS4;1, encoding a cytosolic cysteine desulfhydrase, and BSAS1;1, encoding a cytosolic O-acetylserine sulphydrylase were most highly expressed in both species. This was followed by BSAS3;1 encoding a plastidic β-cyanoalanine synthase which was more highly expressed by 10-fold in P. vulgaris. The data identify BSAS3;1 as a candidate enzyme for the biosynthesis of S-methylcysteine through the use of methanethiol as substrate instead of cyanide. Expression of GLC1 would provide a complete sequence leading to the biosynthesis of γ-Glutamyl-S-methylcysteine in plastids. The detection of S-methylhomoglutathione in P. vulgaris suggested that homoglutathione synthetase may accept, to some extent, γ-Glutamyl-S-methylcysteine as substrate, which might lead to the formation of S-methylated phytochelatins. In conclusion, 454 sequencing was effective at revealing differences in the expression of sulfur metabolic genes, providing information on candidate genes for the biosynthesis of distinct sulfur amino acid γ-Glutamyl dipeptides between P. vulgaris and V. mungo.
普通菜豆(Phaseolus vulgaris)和黑眼豆(Vigna mungo)分别在种子中积累γ-谷氨酰-S-甲基半胱氨酸和γ-谷氨酰-蛋氨酸。在与这些代谢物开始积累的相似发育阶段,通过 454 焦磷酸测序数据对转录本进行了分析。表达序列标签被组装成 Unigenes,并被分配到普通菜豆早期染色体组装的特定基因中。涉及多种硫代谢过程的基因在两种物种中均有表达。Sultr3 成员的表达在普通菜豆中占优势,而 Sultr5 成员的表达在黑眼豆中占优势。细胞质 SERAT1;1 和 -1;2 的表达在普通菜豆中约高出四倍,而质体 SERAT2;1 的表达在黑眼豆中高出两倍。在 BSAS 家族成员中,编码细胞质半胱氨酸脱硫酶的 BSAS4;1 和编码细胞质 O-乙酰丝氨酸硫醇酶的 BSAS1;1 在两种物种中表达水平最高。紧随其后的是编码质体β-氰基丙氨酸合酶的 BSAS3;1,在普通菜豆中表达高出 10 倍。该数据表明,BSAS3;1 可以作为通过使用甲硫醇作为底物而不是氰化物来合成 S-甲基半胱氨酸的候选酶。GLC1 的表达将在质体中提供导致 γ-谷氨酰-S-甲基半胱氨酸生物合成的完整序列。在普通菜豆中检测到 S-甲基同型谷胱甘肽表明,同型谷胱甘肽合酶可能在一定程度上接受 γ-谷氨酰-S-甲基半胱氨酸作为底物,这可能导致 S-甲基化的植物螯合肽的形成。总之,454 测序有效地揭示了硫代谢基因表达的差异,为普通菜豆和黑眼豆中不同硫氨基酸 γ-谷氨酰二肽生物合成的候选基因提供了信息。