八个抗稻瘟病基因在华南籼型杂交水稻中的分布

doi:10.16819/j.1001-7216.2017.6114 299

摘要/Abstract

摘要：

【目的】已克隆的稻瘟病抗性基因Pi1、Pik-p、Pik-h、Pi2、Pi9、Piz-t、Pita、Pii对不同稻区的稻瘟病菌表现较广谱的抗性,被广泛应用于水稻抗瘟性育种。为了明确上述抗性基因在华南稻区杂交稻组合中的分布及其组合的抗病有效性,【方法】利用上述8个抗病基因的功能标记,对华南328个杂交稻组合进行了抗瘟基因型分子检测。【结果】抗性基因Pita和Pii分布频率最高,在测试组合中检出率分别为84.76%与67.68%;其次是Pi2与Pik-p,分别为22.87%与13.72%;检出频率较低的是Pi1、Piz-t和Pik-h,分别为5.18%、3.35%与2.13%,检测的品种都不携带抗性基因Pi9。在单个杂交稻组合中,检出的抗瘟基因数量最多是4个。抗病性评价结果表明,杂交稻组合中检出的抗病基因数量越多,其表现为抗病品种的频率就越高;含4个抗病基因的杂交稻组合中,抗病品种所占比率达91.67%。含有不同抗瘟基因的组合表现出不同水平的抗瘟性,其中Pi2与Pi1对华南稻区稻瘟病的抗病性贡献最大,其他抗病基因的贡献大小依次是Pik-h、Pik-p、Pita、Pii与Piz-t。【结论】本研究为华南稻区杂交稻抗病品种基因型的合理布局以及抗瘟基因的应用提供了科学依据。

关键词: 杂交稻组合, 抗病基因, 稻瘟病, 基因型分析, 分子标记

Abstract:

【Objective】The cloned rice blast resistant genes, Pi1, Pik-p, Pik-h, Pi2, Pi9, Piz-t, Pita and Pii, which showed broad-spectrum blast resistance in different rice growing regions, have been widely used in rice breeding for blast resistance. To clarify the distribution of these genes in hybrid rice in South China, 【Method】the genotypes of resistance genes Pi1, Pik-p, Pik-h, Pi2, Pi9, Piz-t, Pita and Pii in 328 hybrid rice combinations were detected with functional markers. 【Result】The distribution frequency of Pita and Pii was the highest, reaching 84.76% and 67.68% in the tested rice combinations, respectively; followed by Pi2 and Pik-p with the proportion of 22.87% and 13.72%, respectively; Pi1, Piz-t and Pik-h had the lowest distribution frequency, accounting for 5.18%, 3.35% and 2.13% of total; all of these varieties didn’t carry resistance gene Pi9. The number of blast resistance genes detected in hybrid rice combinations were at most four. The resistance evaluation showed that the frequency of resistant varieties rose with increasing number of resistance genes in hybrid combinations. In a hybrid combinations with four resistance genes, the distribution frequency of resistant varieties was 91.67%. Blast resistance contribution assay of the eight resistance genes revealed that Pi2 and Pi1 displayed the greatest resistance contribution in the local rice in South China, followed by Pik-h, Pik-p, Pita, Pii and Piz-t. 【Conclusion】This study provided a scientific basis for the rational distribution of resistant varieties with different genotypes in South China.

Key words: hybrid rice combinations, resistance gene, rice blast, genotype analysis, molecular marker

中图分类号:

汪文娟, 周继勇, 汪聪颖, 苏菁, 封金奇, 陈炳, 冯爱卿, 杨健源, 陈深, 朱小源. 八个抗稻瘟病基因在华南籼型杂交水稻中的分布[J]. 中国水稻科学, 2017, 31(3): 299-306.

Wenjuan WANG, Jiyong ZHOU, Congying WANG, Jing SU, Jinqi FENG, Bing CHEN, Aiqing FENG, Jianyuan YANG, Shen CHEN, Xiaoyuan ZHU. Distribution of Eight Rice Blast Resistance Genes in indica Hybrid Rice in China[J]. Chinese Journal OF Rice Science, 2017, 31(3): 299-306.

图/表 5

表1 各基因特异标记检测引物

Table 1 Specific marker detecting primers for each gene.

引物名称	引物序列(5’-3’)		预期片段大小	标记类型		内切酶		参考文献
Primer name	Primer sequence (5’-3’)	Expected size/bp			Marker type	Enzyme		Reference
Pi2-SNP	F: TACTCTTCGTTGTATAGGAC		462	CAPS		HinfⅠ	[24]
	R: GGAGGAGGAGATGAAATAGAATC
Pi9-SNP	F: CGCCGGTTGATAAGTAAAAGCT		126	dCAPS		Hind Ⅲ	[24]
	R: CAAGAACTAATATCTACCCATGG
Piz-t P/A	F: ATGTGGATGCTGTGTTAT		176	Presence/Absence		—	[24]
	R: TAGTTTGCTGCTCAATAAGTA
K1	F: GCAAGATCAGTACCATCACGAGTAATAGCA		1600	Presence/Absence		—	[19]
	R: GAGTAGCACTCAACGCAAAAGGGCATCGGC
Pik-p2/Pik-h2 SNP	F: CGTGGAAGTTCAACAAAAGG		114	dCAPS		NdeⅠ	[19]
	R: CAGCACCTGTATTATCCCAT
Pik-h1/Pik-p1 SNP	F: TGCTGCAGGTCAGCCAAGCTTA		138	dCAPS		MseⅠ	[19]
	R: CAACCGTTGTTTTGCCTCC
Pi1-SNP	F: CAATAGTCCAGCTAAAACGG		179	CAPS		RsaⅠ	[20]
	R: CATTGCGCCTTTACCTTGT
Pita-SNP	F: GCCGTGGCTTCTATCTTCAGCT		181	dCAPS		PvuⅡ	未发表 Unpublished
	R: GTAACTGTAAAATGACAAGAAAC						未发表 Unpublished
Pii-4SNP	F: TCCAATGCTTCTGAAAGGTAGC		350(240/113)	CAPS		PvuⅡ	[23]
	R: TGGAAACATGAACCCATATCC

图1 基于特异性SNP分子标记的水稻品种抗瘟基因检测 M–DL500。A–区分Pik-p和Pik-h的SNP标记对部分组合的分子检测,CK1–IRBLkp-K60 ( Pik-p+),CK2–IRBLkh-K3 (Pik-h+);B–Pii基因特异性分子标记对部分组合的分子检测,CK1–IRBLi-F5（Pii +）;C–Pi2基因特异性分子标记对部分组合的分子检测,CK1–C101A51 (Pi2+);D–Pita基因特异性分子标记对部分组合的分子检测,CK1–IRBLta-CP1 (Pita+)。泳道1~15:部分杂交稻组合,依次是天优613、泰丰优806、元丰优401、新优1003、裕优132、恒丰优华占、裕优9822、泰丰优2213、盛优华占、信两优3号、乐两优1173、天优华占、恒丰优9802、五丰优1173 与 Y两优1173。

Fig. 1. Detection of R genes in rice cultivars with specific SNP markers. M, DL500. A, Identification of Pik-p and Pik-h with specific SNP marker; CK1, IRBLkp-K60(Pik-p+); CK2, IRBLkh-K3(Pik-h+). B, Identification of Pii with specific SNP marker; CK1, IRBLi-F5(Pii+). C, Identification of Pi2 with specific SNP marker; CK1, C101A51(Pi2+). D, Identification of Pita with specific SNP marker; CK1, IRBLta-CP1 (Pita+). Lanes 1-15, Some of hybrid rice combinations, Tianyou 613, Taifengyou 806, Yuanfengyou 401, Xinyou 1003, Yuyou 132, Hengfengyouhuazhan, Yuyou 9822, Taifengyou 2213, Shengyouhuazhan, Xinliangyou 3, Leliangyou 1173, Tianyouhuazhan, Hengfengyou 9802, Wufengyou 1173, Y liangyou 1173.

图2 抗稻瘟病基因在杂交稻组合中的分布

Fig. 2. Distribution of the blast resistance genes in the hybrid rice combinations.

图3 单个杂交稻组合所含基因数及其所占比率

Fig. 3. Number of blast resistance genes detected in each hybrid rice combination and the percentage.

图4 328个华南稻区杂交稻组合抗性水平分布

Fig. 4. Percentage of resestance levels in the 328 hybrid rice combinations.

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