Mutations in Argonaute5 Illuminate Epistatic Interactions of the K1 and / Loci Leading to Saddle Seed Color Patterns in Glycine max
Mutations in Argonaute5 Illuminate Epistatic Interactions of the K1 and / Loci Leading to Saddle Seed Color Patterns in Glycine max
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DOI：
10.1105/tpc.17.00162
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发表时间：
2017-04-01
期刊：
影响因子：
11.6
通讯作者：
Vodkin, Lila O.
中科院分区：
文献类型：
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作者：
Cho, Young B.;Jones, Sarah I.;Vodkin, Lila O.
大豆（Glycine max）种皮具有独特的、遗传编程的色素沉着模式，隐性k1突变可以上位性地克服显性I和I (I)等位基因，这些等位基因通过产生靶向查尔酮合成酶（CHS） mrna的小干扰rna （sirna）来抑制种子颜色。对未成熟种皮解剖区域的小RNA测序表明，CHS siRNA水平导致i位点的i(i)和i(k)等位基因产生的模式，它们分别将色素限制在种皮的门区或鞍区。为了确定K1位点，我们比较了两条Clark等号染色体的解剖区域的RNA-seq数据，这两条等号染色体具有相似的鞍型，由CHS sirna介导，但不同的基因型(纯合子i（k) K1与纯合子i(i) K1）。通过检测差异表达基因、定位信息和基因组重测序，我们在Glyma. 11G190900中发现了一个129 bp的缺失，该缺失编码Argonaute5 (AGO5)，这是Argonaute家族的成员。对来自不同遗传背景的几个独立鞍型突变体进行扩增子测序，发现影响AGO5的独立病变，从而确定Glyma. 11G190900为K1位点。来自k1等位基因的无功能AGO5导致CHS sirna分布的改变，从而解释了k1突变如何将种皮区域的表型从黄色逆转为色素，即使在正常显性I或I (I)等位基因存在的情况下。
The soybean (Glycine max) seed coat has distinctive, genetically programmed patterns of pigmentation, and the recessive k1 mutation can epistatically overcome the dominant I and i(i) alleles, which inhibit seed color by producing small interfering RNAs (siRNAs) targeting chalcone synthase (CHS) mRNAs. Small RNA sequencing of dissected regions of immature seed coats demonstrated that CHS siRNA levels cause the patterns produced by the i(i) and i(k) alleles of the I locus, which restrict pigment to the hilum or saddle region of the seed coat, respectively. To identify the K1 locus, we compared RNA-seq data from dissected regions of two Clark isolines having similar saddle phenotypes mediated by CHS siRNAs but different genotypes (homozygous i(k) K1 versus homozygous i(i) k1). By examining differentially expressed genes, mapping information, and genome resequencing, we identified a 129-bp deletion in Glyma. 11G190900 encoding Argonaute5 (AGO5), a member of the Argonaute family. Amplicon sequencing of several independent saddle pattern mutants from different genetic backgrounds revealed independent lesions affecting AGO5, thus establishing Glyma. 11G190900 as the K1 locus. Nonfunctional AGO5 from k1 alleles leads to altered distributions of CHS siRNAs, thus explaining how the k1 mutation reverses the phenotype of the seed coat regions from yellow to pigmented, even in the presence of the normally dominant I or i(i) alleles.