水稻对穗枯病的抗病机理初步研究

doi:10.16819/j.1001-7216.2017.6167 551

摘要/Abstract

摘要：

目的水稻在孕穗期比苗期更容易感染穗枯病(Bacterial panicle blight of rice)并出现病症。本研究旨在探究水稻不同时期对穗枯病的抗性机理,为培育抗病水稻品种奠定基础。方法采用喷雾法和注射法分别对苗期和孕穗期的抗、感病水稻品种接种颖壳伯克氏菌（Burkholderia glumae）,测定处理组与对照组的3种抗氧化酶(POD、CAT、SOD)活性的差异,并利用实时荧光定量PCR测定5种防卫反应基因(PR1a、PR10b、Rcht、LOX、PAL)的表达量。结果 B. glumae的侵染能引起水稻活性氧的积累,提高抗氧化酶活性,使部分防卫反应基因大量表达,但是苗期和孕穗期的应答有较大区别。水稻孕穗期的抗氧化酶(SOD、CAT和POD)活性和防卫反应基因(PR10b、Rcht和PAL)的表达量比苗期高,但PR1a和LOX的表达量却低于苗期。结论 B. glumae能诱导孕穗期的水稻产生更多抗病反应,参与抗病反应的主要是水杨酸信号传导途径。

关键词: 水稻, 穗枯病, 抗氧化酶, 防卫反应基因, 实时荧光定量PCR

Abstract:

【Objective】Rice is more susceptible to bacterial panicle blight at the booting stage than seedling stage. The objective of the study is to understand the resistant mechanism in different growing stage of rice and lay a basis for breeding rice varieties with high resistance.【Method】The spraying and injection method were used to inoculate seedlings and panicles of resistant and susceptible rice with Burkholderia glumae, respectively. The activities of three antioxidant enzymes (peroxidase, catalase and superoxide dismutase) of treatments and control were investigated. Real-time PCR was used to detect the expression of five defense response genes (PR1a, PR10b, Rcht, LOX and PAL).【Result】The infection of B. glumae induced the accumulation of reactive oxygen species, the increase of antioxidant enzymes activities and the higher expression of some defense genes. However, the defense response was different at the seedling stage and booting stage of rice. At the booting stage, the activities of antioxidant enzymes (peroxidase, catalase and superoxide dismutase) and the expression of PR10b, Rcht and PAL were higher, but the expression levels of PR1a and LOX were lower than those at the seedling stage.【Conclusion】B. glumae could induce more defense response of rice in booting stage, salicylic acid signaling pathway played an important role during the defense.

Key words: rice, bacterial panicle blight, antioxidant enzyme, defense gene, real-time fluorescence quantitative PCR

中图分类号:

S435.111.4⁺5

李路, 徐以华, 梁梦琦, 王玲, 刘连盟, 侯雨萱, 黎起秦, 黄世文. 水稻对穗枯病的抗病机理初步研究[J]. 中国水稻科学, 2017, 31(5): 551-558.

Lu LI, Yihua XU, Mengqi LIANG, Ling WANG, Lianmeng LIU, Yuxuan HOU, Qiqin LI, Shiwen HUANG. A Primary Study on the Mechanism Behind Resistance to Bacterial Panicle Blight of Rice[J]. Chinese Journal OF Rice Science, 2017, 31(5): 551-558.

图/表 9

表1 荧光定量PCR的引物

Table 1 Specific primers for real-time PCR

基因 Gene	登录号 Accession No.	引物序列 Primer sequence	扩增长度 Length/bp
PR1a	CA758799.1	F:GTTATCCTGCTGCTTGCTG R:GTACCACTGCTTCTCCGACACCCAC	299
PR10b	CA755132.1	F: TGAGGTGTTGGAGGTTAAGAG R:AGGGTGAGCGACGAGGTA	217
Rcht	AB016497.1	F:CGCCGTCATCCAGAACCAG R:AGATAAACAAGGCGACTTCTCCAC	179
LOX	FJ607153	F:CGCCATCATCGTCACCAA R: CGTCTGTCCCGGCAAGTAA	199
PAL	EF576408	F:TTCCCGCTCTACCGCTTCGT R:GCTCGCCGTTCCACTCCTTG	193
Ubq	AY072820.1	F: CTCGCCGACTACAACATC R: GACATCAGCACCACAAATAC	147

图1 不同抗性水稻品种中过氧化物酶(POD)的活性

Fig. 1. Activity of peroxidase(POD) in rice varieties with different resistance to bacterial panicle blight.

图2 不同抗性水稻品种中过氧化氢酶(CAT)的活性

Fig. 2. Activity of catalase(CAT) in rice varieties with different resistance to bacterial panicle blight.

图3 不同抗性水稻品种中超氧化物歧化酶(SOD)的活性

Fig. 3. Activity of superoxide dismutase(SOD) in rice varieties with different resistance to bacterial panicle blight.

图4 不同抗性水稻品种中PR1a相对表达量

Fig. 4. Relative expression of PR1a in rice varieties with different resistance to bacterial panicle blight.

图5 不同抗性水稻品种中PR10b相对表达量

Fig. 5. Relative expression of PR10b in rice varieties with different resistance to bacterial panicle blight.

图6 不同抗性水稻品种中Rcht相对表达量

Fig. 6. Relative expression of Rcht in rice varieties with different resistance to bacterial panicle blight.

图7 不同抗性水稻品种中LOX相对表达量

Fig. 7. Relative expression of LOX in rice varieties with different resistance to bacterial panicle blight.

图8 不同抗性水稻品种中PAL相对表达量

Fig. 8. Relative expression of PAL in rice varieties with different resistance to bacterial panicle blight.

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