Resistance to Early Senescence and Rice Blast/Blight: A Mutant Variety and Expressions of Responsible Genes
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摘要:
目的 叶片早衰影响水稻的产量和品质，早衰突变体是研究水稻衰老机制的良好载体，鉴定、分析早衰突变体的表型特征，有助于了解突变体的遗传规律，为相关基因的克隆和功能研究奠定基础。 方法 人工接种鉴定了早衰突变体w14的稻瘟病和水稻白叶枯病抗性，同时通过对离体叶片的黑暗诱导和叶绿素含量测定，鉴定了突变体w14的早衰表型；再分别对早衰突变体w14和野生型粳稻品种云引（简称‘YY’）中水稻早衰相关基因和病程相关基因的表达进行分析，研究早衰突变体w14的早衰类型及与野生型在防御系统上的差异。 结果 黑暗诱导处理24 h后，突变体w14的叶绿素含量显著低于野生型YY（P＜0.05）。黑暗诱导处理48 h以上，突变体w14的叶绿素含量极显著低于野生型YY（P＜0.01）。突变体w14中衰老相关基因SGR、Osh36、Osh69、PAO、NYC3和RCCR1的表达均显著高于野生型YY。分别接种水稻白叶枯和稻瘟病后，突变体w14均表现为比野生型YY更感病，且病程相关基因PR1a、PR4、Cth1、PR1b、PBZ1、PR3等的表达在突变体中显著上调。 结论 突变体w14具有典型的衰老和感稻瘟病、水稻白叶枯病的表型，与野生型相比，在突变体中与病程相关基因的表达发生了显著改变，即突变体防御系统的改变导致其出现感病的表型。 Abstract:Objective A mutant rice showing signs of early senescence on leaves and susceptibility to rice blast and blight was used for phenotypic and genetic analyses to study the genes associated with the aging mechanism and vulnerability to the diseases of rice plants. Method Resistance to blast and bacterial blight of the mutant rice w14 with symptoms of early senescence was tested by artificial inoculation. Phenotype classification of the mutant was determined by chlorophyll content measured in vitro under induced darkness. Expressions of the genes related to senescence and pathogenesis in the mutant and wild-type japonica rice variety, Yunyin (YY), were compared. Result After inducing darkness for 24h, the chlorophyll content in w14 was 5% lower than that in YY, and the difference reached a 1% statistical significance level in 48 h. The expressions of senescence-related genes SGR, Osh36, Osh69, PAO, NYC3, and RCCR1 in w14 were significantly higher than those in the wild-type counterpart. The mutant rice was more susceptible to the artificial inoculations of rice blast and bacterial blight, and the pathogenesis-related genes PR1a, PR4, Cth1, PR1b, PBZ1, and PR3 significantly up-regulated in w14 than in YY. Conclusion The mutant rice w14 showed the typical senescence phenotypes with a higher susceptibility to rice blast and bacterial blight than the wild-type YY. The expressions of pathogenesis-related genes in the mutants were significantly altered that substantially affected its defense system leading to the early senescence and disease susceptibility as observed in the study. -
Key words:
- Rice /
- mutant /
- early senescence /
- chlorophyll /
- resistance /
- gene expression
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图 1 突变体w14和野生型的离体叶片黑暗诱导处理鉴定结果
注：a：处理0～96 h离体叶片的衰老表型；b：处理0～96 h离体叶片的叶绿素含量测定；Chla、Chlb和Chlx分别代表叶绿素a、叶绿素b和类胡萝卜素；*和**分别表示在0.05和0.01水平差异显著。
Figure 1. Early senescence on mutant w14 and wild-type YY rice plants after in vitro darkness-induction
Note：a: Senescence phenotype of leaves treated for 0-96 h. b: Chlorophyll content of leaves treated for 0-96 h; *and** show significant difference at 0.05 and 0.01 levels, respectively.
图 2 突变体w14与野生型YY接种水稻白叶枯病发病后的症状
注：a：成株期田间接种水稻白叶枯病后的发病症状；b：室内苗期接种和大田成株期接种后发病结果；**表示在0.01水平显著差异。
Figure 2. Symptoms on diseased mutant w14 and wild-type YY after inoculation of rice bacterial blight
Note：a: Symptoms of rice bacterial blight after field inoculation on rice plants at adult stage. b: Statistics on occurrence of bacterial blight on rice plants after indoor inoculation at seedling stage and field inoculation at adult stage; ** shows significant difference at 0.01 level.
图 3 突变体w14与野生型YY与衰老相关基因的表达
注：*和**分别表示在0.05和0.01水平显著差异。
Figure 3. Expressions of senescence-related genes of mutant w14 and wild-type YY
Note：* and ** show significant difference at 0.05 and 0.01 levels, respectively.
图 4 突变体w14与野生型YY水稻病程相关基因的表达分析
注：a：CEBip、PR1b、JAR1、PAL2、PR5和PBZ1的表达；b：PR1a、PAL1、PR3、PR4和Cth1的表达；YY-s、YY-m、w14-s、w14-m中的s和m分别表示苗期和成熟期。
Figure 4. Expressions of rice pathogenesis-related genes of mutant w14 and wild-type YY
Note：a: Expression levels of CEBip, PR1b, JAR1, PAL2, PR5, and PBZ1. b: Expression levels of PR1a, PAL1, PR3, PR4, and Cth1; s and m in YY-s, YY-m, w14-s and w14-m represent seedling stage and maturity stage, respectively.
表 1 突变体w14和野生型的稻瘟病抗性鉴定结果
Table 1. Resistance to blast disease of mutant w14 and wild-type YY
菌株 Strain 表型 Phenotype（w14/YY/LTH） 菌株 Strain 表型 Phenotype（w14/YY/LTH） SM17021-2 S/S/S NH15092 S/S/S NH15039 S/S/S NH15040 S/R/S JY14027 S/S/S NH17025 R/R/S SM17019-2 S/R/S JY15013 S/S/S NH17026-8 S/S/S SM17019-1 R/R/S NH13093 S/R/S JY15027 R/R/S JL15140 R/R/S SM17020-7 R/R/S JL15131 R/R/S NJ08022 S/R/S SH17004 S/R/S NH17029 R/R/S SM17021 S/S/S SM17020-3 S/S/S NH17028 S/R/S NH17026-4 S/R/S JY15015 S/R/S NH17026-7 R/R/S 
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