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Virus-induced gene silencing in soybean seeds and the emergence stage of soybean plants with Apple latent spherical virus vectors

机译:苹果潜伏球形病毒载体在大豆种子中的病毒诱导基因沉默和大豆植株的出苗阶段

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Virus-induced gene silencing (VIGS) has great potential as a reverse-genetics tool in plant genomics. In this study, we examined the potential of VIGS in soybean seeds and the emergence stage of soybean plants using Apple latent spherical virus (ALSV) vectors. Inoculation of an ALSV vector (soyPDS-ALSV) carrying a fragment of the soybean phytoene desaturase (soyPDS) gene into soybean seedlings resulted in a highly uniform photo-bleached phenotype, typical of PDS inhibition, on the upper leaves throughout plant growth. The photo-bleached phenotype was also found on all immature pods, all seed coats, and about 50% embryos of seeds on soybean plants infected with soyPDS-ALSV. Infection with an ALSV vector (soyIFS2-ALSV) having a fragment of soybean isoflavone synthase 2 (soyIFS2) gene also led to a reduction of the levels of both soyIFS2- and soyIFS1- mRNAs and an isoflavone content in the cotyledons of about 36% mature seeds of infected soybean plants. Furthermore, VIGS of soyPDS was induced in the next generation plants by the seed transmission of soyPDS-ALSV. Thus ALSV vectors will be useful for studying gene functions in the reproductive stages and early growth stages, such as emergence and cotyledon stages, in addition to the vegetative stages of soybean plants.
机译:病毒诱导的基因沉默(VIGS)作为植物基因组学中的逆向遗传工具具有巨大潜力。在这项研究中,我们使用苹果潜伏球形病毒(ALSV)载体检查了VIGS在大豆种子中的潜力以及大豆植物的出苗阶段。将携带大豆八氢番茄红素去饱和酶(soyPDS)基因片段的ALSV载体(soyPDS-ALSV)接种到大豆幼苗中,会在整个植物生长的上部叶片上产生高度均一的光漂白表型,典型的是PDS抑制。在感染了soyPDS-ALSV的大豆植株上,所有未成熟的豆荚,所有种皮和大约50%的种子胚上也发现了光漂白的表型。感染具有大豆异黄酮合酶2(soyIFS2)基因片段的ALSV载体(soyIFS2-ALSV)也会导致soyIFS2-和soyIFS1- mRNA的水平降低,并且子叶中异黄酮的含量约为36%成熟大豆植物的种子。此外,在下一代植物中,通过soyPDS-ALSV的种子传播诱导了soyPDS的VIGS。因此,除了大豆植物的营养阶段外,ALSV载体还将用于研究生殖阶段和早期生长阶段(例如出苗和子叶阶段)的基因功能。

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