Improving Rice Blast Resistance by Mining Broad-Spectrum Resistance Genes at Pik Locus

doi:10.1016/j.rsci.2022.01.002

Abstract

Abstract:

Magnaporthe oryzae is known for its genetic diversity and pathogenic variability, leading to rapid breakdown of resistance in rice. Incorporating multiple broad-spectrum blast resistance genes into rice cultivars would extend disease resistance longevity. Effective resistance breeding in rice therefore requires continual enrichment of the reservoir of resistance genes and alleles. We conducted a large-scale screen of rice blast resistance in about 2 000 rice accessions. Among them, 247 accessions showed at least medium resistance to the natural infection of rice blast and 7 novel Pik alleles were identified from them. Variations in gene sequences were then correlated with the phenotypic trait to enable the identification of favorable alleles. Among the seven novel Pik alleles, the resistant rate of Pik-R0/ME/7017 donors was greater than 80%, and the disease score was less than 3. Through molecular marker-assisted backcross breeding, we successfully transferred the three Pik alleles, Pik-R0/ME/7017, into an elite cultivated line Kongyu 131 to obtain BC₃F₂ lines, which showed enhanced resistance to rice blast compared with the recurrent parent. Assessment of these near-isogenic lines in the greenhouse using 31 isolates of M. oryzae from Heilongjiang Province of China revealed that the resistant levels of the BC₃F₂ lines with Pik-R0/ME/7017 were significantly higher than those of the established cloned resistance genes Pik-m and Pi1. Exploring such alleles will enrich our gene library for resistance to rice blast.

Key words: genetic diversity, near-isogenic line, Pik gene, resistance gene allele, rice blast, R gene

Zhou Ying, Wan Tao, Yuan Bin, Lei Fang, Chen Meijuan, Wang Qiong, Huang Ping, Kou Shuyan, Qiu Wenxiu, Liu Li. Improving Rice Blast Resistance by Mining Broad-Spectrum Resistance Genes at Pik Locus[J]. Rice Science, 2022, 29(2): 133-142.

Figures/Tables 6

References 42

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Pik allele	Accession ^a	No. of accessions carrying the allele	Identity to Pik-m (%)	No. of SNPs			No. of InDels
Pik allele	Accession ^a	No. of accessions carrying the allele	Identity to Pik-m (%)	Total	Pik-1	Pik-2	Total	Pik-1	Pik-2
Pik-m	Tsuyuake	13	‒	‒	‒	‒	‒	‒	‒
Pik-s	Khao Phoi	15	99.97	4	3	0	0	0	0
Pi1	Tetep	4	99.91	12	3	1	5	5	0
Pik-NS	Nsicrc 122	19	99.85	23	8	6	7	5	0
Pik-R0	R03138	10	98.34	230	20	6	200	6	0
Pik-ME	Meixiangzhan	1	97.17	389	116	9	257	53	0
Pik-CE	IR65482-4-136-2-2-B	1	97.31	373	112	7	246	55	0
Pik-7017	Red Khosha Cerma	4	97.31	372	105	8	246	49	0
Pik-YD	Yangdao 4038	49	97.16	390	113	9	259	62	0
Pik-ZH	Zhouhui 338	3	98.28	232	15	8	201	4	0

Pik allele	Accession ^a	No. of accessions carrying the allele	Identity to Pik-m (%)	No. of SNPs			No. of InDels
Pik allele	Accession ^a	No. of accessions carrying the allele	Identity to Pik-m (%)	Total	Pik-1	Pik-2	Total	Pik-1	Pik-2
Pik-m	Tsuyuake	13	‒	‒	‒	‒	‒	‒	‒
Pik-s	Khao Phoi	15	99.97	4	3	0	0	0	0
Pi1	Tetep	4	99.91	12	3	1	5	5	0
Pik-NS	Nsicrc 122	19	99.85	23	8	6	7	5	0
Pik-R0	R03138	10	98.34	230	20	6	200	6	0
Pik-ME	Meixiangzhan	1	97.17	389	116	9	257	53	0
Pik-CE	IR65482-4-136-2-2-B	1	97.31	373	112	7	246	55	0
Pik-7017	Red Khosha Cerma	4	97.31	372	105	8	246	49	0
Pik-YD	Yangdao 4038	49	97.16	390	113	9	259	62	0
Pik-ZH	Zhouhui 338	3	98.28	232	15	8	201	4	0

Pik allele protein	Accession^a	No. of amino acids^b		Identity to Pik-m protein (%)		No. of different amino acids			No. of InDels
Pik allele protein	Accession^a	Pik-1	Pik-2	Pik-1	Pik-2	Total	Pik-1	Pik-2	Total	Pik-1	Pik-2
Pik-m	Tsuyuake	1 143	1 021	‒	‒	‒	‒	‒	‒	‒	‒
Pik-s	Khao Phoi	1 143	1 021	99.83	100.00	2	2	0	0	0	0
Pi1	Tetep	1 143	1 021	99.91	99.90	1	1	0	0	0	0
Pik-NS	Nsicrc 122	1 143	1 021	99.74	99.80	5	3	2	0	0	0
Pik-R0	R03138	1 143	1 021	99.13	99.80	8	6	2	0	0	0
Pik-ME	Meixiangzhan	1 146	1 021	94.59	99.51	67	62	5	3	3	0
Pik-CE	IR65482-4-136-2-2-B	1 142	1 021	95.01	99.71	60	57	3	1	1	0
Pik-7017	Red Khosha Cerma	1 142	1 021	95.10	99.61	60	56	4	1	1	0
Pik-YD	Yangdao 4038	1 144	1 021	94.85	99.51	64	59	5	1	1	0
Pik-ZH	Zhouhui 338	1 143	1 021	99.21	99.61	13	9	4	0	0	0

Pik allele protein	Accession^a	No. of amino acids^b		Identity to Pik-m protein (%)		No. of different amino acids			No. of InDels
Pik allele protein	Accession^a	Pik-1	Pik-2	Pik-1	Pik-2	Total	Pik-1	Pik-2	Total	Pik-1	Pik-2
Pik-m	Tsuyuake	1 143	1 021	‒	‒	‒	‒	‒	‒	‒	‒
Pik-s	Khao Phoi	1 143	1 021	99.83	100.00	2	2	0	0	0	0
Pi1	Tetep	1 143	1 021	99.91	99.90	1	1	0	0	0	0
Pik-NS	Nsicrc 122	1 143	1 021	99.74	99.80	5	3	2	0	0	0
Pik-R0	R03138	1 143	1 021	99.13	99.80	8	6	2	0	0	0
Pik-ME	Meixiangzhan	1 146	1 021	94.59	99.51	67	62	5	3	3	0
Pik-CE	IR65482-4-136-2-2-B	1 142	1 021	95.01	99.71	60	57	3	1	1	0
Pik-7017	Red Khosha Cerma	1 142	1 021	95.10	99.61	60	56	4	1	1	0
Pik-YD	Yangdao 4038	1 144	1 021	94.85	99.51	64	59	5	1	1	0
Pik-ZH	Zhouhui 338	1 143	1 021	99.21	99.61	13	9	4	0	0	0

Gene	Accession ^a	Enshi		Yichang		Jiamusi
Gene	Accession ^a	2018	2019	2018	2019	2018
CK1	Lijiangxintuanheigu	S	S	HS	S	HS
CK2	Jin 23B	HS	HS	HS	HS	S
Pik-m	R107	HS	HS	HS	HS	MR
Pik-s	Jiahua 1	R	HR	HR	HR	MR
Pi1	Tetep	HR	HR	HR	HR	S
Pik-NS	Nsicrc 122	HR	HR	HR	HR	MR
Pik-R0	R03138	HR	HR	HR	HR	MR
Pik-ME	Meixiangzhan	HR	HR	MR	MR	MR
Pik-CE	IR65482-4-136-2-2-B	S	MR	HR	MR	MR
Pik-7017	Red Khosha Cerma	MS	HR	MS	MR	MS
Pik-YD	Yangdao 4038	R	HR	HR	HR	R
Pik-ZH	Zhouhui 338	HS	HS	HS	HS	MR