Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (23): 4933-4942.doi: 10.3864/j.issn.0578-1752.2021.23.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Overexpression of OsPR1A Enhanced Xa21-Mediated Resistance to Rice Bacterial Blight

LIU YuQing1(),YAN GaoWei1,ZHANG Tong2,LAN JinPing3,GUO YaLu4,LI LiYun1,LIU GuoZhen1(),DOU ShiJuan1()   

  1. 1College 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
  • Received:2021-05-08 Accepted:2021-07-02 Online:2021-12-01 Published:2021-12-06
  • Contact: GuoZhen LIU,ShiJuan DOU E-mail:liuyuqing_mbb@126.com;gzhliu@hebau.edu.cn;dsj75@126.com

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.

Key words: rice, Xa21, bacterial blight, OsPR1A, vector construction, western blot

Fig. 1

The construction and verification of transformation plasmid for OsPR1a-OX"

Fig. 2

The identification of OsPR1a overexpression transgenic plants"

Fig. 3

The 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"

Fig. 4

The 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)"

Fig. 5

Protein expression characteristics of OsPR1A overexpression transgenic plants after Xoo inoculation"

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