作物学报 ›› 2024, Vol. 50 ›› Issue (8): 2131-2142.doi: 10.3724/SP.J.1006.2024.44002
李旭娟1,2(), 李纯佳1,2, 田春艳1,2, 孔春艳1,2, 徐超华1,2, 刘新龙1,2,*(
)
LI Xu-Juan1,2(), LI Chun-Jia1,2, TIAN Chun-Yan1,2, KONG Chun-Yan1,2, XU Chao-Hua1,2, LIU Xin-Long1,2,*(
)
摘要:
甘蔗(Saccharum spp. hybrid)是重要的糖料和能源作物, 分蘖是影响其产量的重要性状之一。硝酸盐转运蛋白1/肽转运蛋白家族(NITRATE TRANSPORTER 1 (NRT1)/PEPTIDE TRANSPORTER (PTR) family, NPF)成员在植生长发育中发挥重要作用, 甘蔗中NPF基因的挖掘利用可为甘蔗品种分蘖遗传调控和产业升级奠定重要基础。本研究通过前期转录组数据结合反转录PCR (reverse transcription PCR, RT-PCR)从甘蔗品种ROC22中获得其NPF家族成员6.4基因的cDNA全长序列(命名为ScNPF6.4), 并对其进行序列结构、理化性质、系统进化等分析, 然后开展了该基因在苗期甘蔗中的组织特异性、甘蔗腋芽萌发过程中的表达情况以及响应激素处理的表达模式分析, 最后将该基因遗传转化水稻过表达进行功能验证。结果表明, ScNPF6.4的cDNA包含1个1806 bp的开放阅读框, 编码601个氨基酸, 属于易化子超家族(major facilitator superfamily, MFS)蛋白, 其蛋白分子量为63.9 kD, 理论等电点为9.23, 包含12个跨膜螺旋区。该蛋白不存在信号肽, 具疏水性, 属于一类稳定的非分泌蛋白, 系统进化分析表明其属于NPF 6.4亚族蛋白。亚细胞定位分析表明该蛋白定位于内质网。组织特异性分析发现, ScNPF6.4在苗期甘蔗根中相对表达量最高, 在叶和茎基部相对表达量较低; ScNPF6.4在不同甘蔗品种腋芽萌发阶段均上调表达; 适量浓度的植物外源激素6-BA、ABA、GA3、IBA、乙烯利和SLs均能诱导苗期甘蔗中ScNPF6.4的上调表达; 水稻中异位过表达ScNPF6.4可增加水稻分蘖数并提前孕穗。以上结果表明, ScNPF6.4正调控甘蔗分蘖芽萌发, 其表达受外源植物激素调控, 该基因过表达可增加水稻分蘖数并促进提前孕穗, 克隆该基因可为甘蔗分蘖早生快发高产育种提供重要的基因资源。
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