Overexpression of OsPR1A Enhanced Xa21-Mediated Resistance to Rice Bacterial Blight
LIU YuQing,1, YAN GaoWei1, ZHANG Tong2, LAN JinPing3, GUO YaLu4, LI LiYun1, LIU GuoZhen,1, DOU ShiJuan,11College of Life Sciences, Hebei Agricultural University, Baoding 071001, Hebei 2Beijing Institute of Biological Products Co., Ltd, Beijing 102600 3Life Science Research Center, Hebei North University, Zhangjiakou 075000, Hebei 4Agricultural Genomics Institute, Chinese Academy of Agricultural Sciences, Shenzhen 518116, Guangdong
Abstract 【Background】Previous studies revealed that the expression of pathogenesis-related protein OsPR1A was regulated by the upstream resistance gene Xa21. The rapid induction of OsPR1A protein at early stage after inoculation was crucial in Xa21-mediated rice bacterial blight resistance. The expression of OsPR1A was induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1A was well known as a marker gene to demonstrate the reaction between rice and pathogen, however, no direct evidence was obtained for the biological function of OsPR1A. 【Objective】 In this paper, transgenic plants overexpression OsPR1A were obtained and the phenotype and agronomic traits were investigated. The relationship between OsPR1A expression and resistance were surveyed to further explore the function of OsPR1A in the process of rice resistance to bacterial blight.【Method】The construct of OsPR1a-OX was transferred to rice recipient 4021 via Agrobacterium-mediated protocol. Positive homozygous transgenic lines were identified by PCR and western blot (WB) respectively. At the mature stage, the phenotype and agronomic traits of OsPR1A overexpression rice plants were investigated, including plant height, spike length, tiller number, seed setting rate and grain size. Rice seedlings of TP309, 4021 and OsPR1A overexpression plants grown for two weeks were inoculated with Xoo at 31℃. The length of lesions was measured at 0, 2, 4, 6, 8, 10, and 12 days post-inoculation (dpi) respectively. At 0, 4 and 6 dpi, the rice leaves of TP309, 4021 and OsPR1A overexpression plants were collected to extract total protein, and the expression profiling of OsPR1A were surveyed by WB.【Result】The OsPR1a-OX transformation vector was constructed and transformed into recipient 4021. Two homozygous OsPR1A overexpression lines (#704 and #709) were identified. At the mature stage, the phenotype and agronomic traits of the OsPR1A overexpression plants were investigated. Compared with the control 4021, #704 and #709 lines showed lower plant height, shorter panicle length, fewer tiller number, lower seed-setting rate. The grain size in transgenic rice plants were larger, which might be related to the lower seed-setting rate. At 31℃, the lesion length of OsPR1A overexpression plants was significantly shorter than that of the control 4021 (P<0.05). At 0, 4, and 6 dpi, the abundance of OsPR1A expression of overexpression plants was higher than that of 4021 and TP309, and the high level of OsPR1A protein might contribute to the resistance of Xoo.【Conclusion】OsPR1A overexpression transgenic plants were obtained by the Agrobacterium-mediated method. Overexpression of OsPR1A affected the normal development of rice plants and also enhanced the resistance to bacterial blight mediated by Xa21. Keywords:rice;Xa21;bacterial blight;OsPR1A;vector construction;western blot
PDF (1495KB)元数据多维度评价相关文章导出EndNote|Ris|Bibtex收藏本文 本文引用格式 刘玉晴, 燕高伟, 张彤, 兰金苹, 郭亚璐, 李莉云, 刘国振, 窦世娟. 超表达OsPR1A增强了Xa21介导的水稻对白叶枯病的抗性反应. 中国农业科学, 2021, 54(23): 4933-4942 doi:10.3864/j.issn.0578-1752.2021.23.001 LIU YuQing, YAN GaoWei, ZHANG Tong, LAN JinPing, GUO YaLu, LI LiYun, LIU GuoZhen, DOU ShiJuan. Overexpression of OsPR1A Enhanced Xa21-Mediated Resistance to Rice Bacterial Blight. Scientia Acricultura Sinica, 2021, 54(23): 4933-4942 doi:10.3864/j.issn.0578-1752.2021.23.001
含有水稻目的片段OsPR1a(LOC_Os07g03710)全长的cDNA序列的质粒购自日本农业生物资源研究所水稻基因组资源中心(Rice Genome Resource Center,National Institute of Agrobiological Sciences)。扩增目的片段使用的引物分别为OsPR1a-OX-F:5’-GCGGTACCATGGCGAGTTCGTCGAGCAGG-3’(下划线为KpnⅠ酶切位点),OsPR1a-OX-R:5’-GCGAATTCTCAGTAGGGAGATTGGCCGAC-3’(下划线为EcoRⅠ酶切位点)。
a:水稻成熟期表型图,bar=100 cm;b:水稻穗部表型图,bar=10 cm;c:水稻籽粒表型图,bar=1cm;d:株高统计学分析;e:穗长统计学分析;f:分蘖数统计学分析;g:结实率统计学分析;*表示P<0.05,**表示P<0.01。下同 Fig. 3The phenotype and agronomic traits of OsPR1a overexpression transgenic plants
a: Phenotype of rice at mature stage, bar=100 cm; b: Phenotype of rice panicle, bar=10 cm; c: Phenotype of rice grain, bar=1 cm; d: Statistical analysis of plant height; e: Statistical analysis of panicle length; f: Statistical analysis of tiller number; g: Statistical analysis of seed setting rate; * means P <0.05, ** means P<0.01. The same as below
a:接种12 d内不同材料的叶片病斑生长曲线,平均值±标准误差(n=4);b:水稻接种Xoo第12天时不同抗感材料和OsPR1a-OX转基因材料的叶片表型图,bar=1 cm;c:接种第12天时不同材料的叶片病斑长度统计学分析,平均值±标准误差(n=4) Fig. 4The lesion growth curve and phenotype of OsPR1a overexpression transgenic plants after inoculation.
a: The growth curve of leaf lesions of different materials within 12 days of inoculation, average ± standard error (n=4); b: Leaf phenotype of TP309, 4021 and OsPR1a-OX transgenic materials at 12 dpi, bar=1 cm; c: Statistical analysis of leaf lesion length of different materials at 12 dpi, average ± standard error (n=4)
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