James M G, Robertson D S, Myers A M
Department of Biochemistry and Biophysics, Iowa State University, Ames 50011, USA.
Plant Cell. 1995 Apr;7(4):417-29. doi: 10.1105/tpc.7.4.417.
In maize kernels, mutations in the gene sugary1 (su1) result in (1) increased sucrose concentration; (2) decreased concentration of amylopectin, the branched component of starch; and (3) accumulation of the highly branched glucopolysaccharide phytoglycogen. To investigate further the mechanisms of storage carbohydrate synthesis in maize, part of the su1 gene locus and a cDNA copy of the su1 transcript were characterized. Five new su1 mutations were isolated in a Mutator background, and the mutant allele su1-R4582::Mu1 was isolated by transposon tagging. The identity of the cloned element as the su1 gene locus was confirmed by the cosegregation of restriction fragment length polymorphisms in the same or nearby genomic intervals with three additional, independent su1 mutations. Pedigree analysis was also used to confirm the identity of su1. A 2.8-kb mRNA that is homologous to the cloned gene was detected in maize kernels, and a 2.7-kb cDNA clone was isolated based on hybridization to the genomic DNA. Specific portions of the cDNA hybridized with multiple segments of the maize genome, suggesting that su1 is part of a multigene family. The cDNA sequence specified a polypeptide of at least 742 amino acids, which is highly similar in amino acid sequence to bacterial enzymes that hydrolyze alpha-(1-->6) glucosyl linkages of starch. Therefore, debranching of glucopolysaccharides is seemingly part of the normal process of starch biosynthesis, and the final degree of branch linkages in starch most likely arises from the combined actions of branching and debranching enzymes.
在玉米籽粒中,含糖1基因(su1)发生突变会导致:(1)蔗糖浓度增加;(2)支链淀粉(淀粉的分支成分)浓度降低;(3)高度分支的葡聚糖植物糖原积累。为了进一步研究玉米中储存碳水化合物合成的机制,对su1基因座的一部分和su1转录本的cDNA拷贝进行了表征。在Mutator背景下分离出5个新的su1突变体,并通过转座子标签法分离出突变等位基因su1-R4582::Mu1。通过同一或附近基因组区间的限制性片段长度多态性与另外3个独立的su1突变体的共分离,证实了克隆元件为su1基因座。系谱分析也用于确认su1的身份。在玉米籽粒中检测到一种与克隆基因同源的2.8 kb mRNA,并基于与基因组DNA的杂交分离出一个2.7 kb的cDNA克隆。cDNA的特定部分与玉米基因组的多个片段杂交,表明su1是一个多基因家族的一部分。cDNA序列编码一个至少742个氨基酸的多肽,其氨基酸序列与水解淀粉α-(1→6) 葡糖基连接的细菌酶高度相似。因此,葡聚糖的去分支似乎是淀粉生物合成正常过程的一部分,淀粉中分支连接的最终程度很可能源于分支酶和去分支酶的共同作用。