Genetic Analysis and Identification of Brown Planthopper Resistance Gene in indica Rice Accession 570011

doi:10.16819/j.1001-7216.2023.220601

Abstract

Abstract:

【Objective】The aim is to identify brown planthopper (BPH) resistance gene in resistant indica rice accession 570011, and provide new materials for insect-resistant rice breeding.【Method】A seedling bulk test was conducted to evaluate the BPH resistance of F₃ population, and a local genetic map was developed based on the genotype of the mapping population by JoinMap 3.0. The resistance locus was then detected and genetic effect was evaluated by MapQTL 5. The expression of one candidate resistance gene was analyzed with qRT-PCR, and the corresponding CDS and amino acid sequences of the candidate gene were compared.【Result】Accession 570011 showed high resistance to BPH at the seedling stage and significant antibiosis and antixenosis against BPH. The ratio of resistant lines (score < 7) to susceptible lines (score≥7) in F₃ population was 89:35, which accorded with the separation law of one dominant gene. One BPH resistance locus was detected in the 39 cM region flanked by markers 4M18675 and 4M24.64 on chromosome 4, which could be allelic to the cloned BPH6. qRT-PCR analysis indicated that the expression of BPH6 allele in accession 570011 was significantly higher than that in susceptible line 9311. Accession 570011 respectively shared the similarity of 99.08% and 97.96% in CDS and amino acid sequence comparing with the cloned gene BPH6. Moreover, they closely clustered in the phylogenetic tree. All the results suggested that the BPH resistance gene derived from accession 570011 was allelic to BPH6.【Conclusion】Accession 570011 is highly resistant to BPH and carries one major resistance gene, which is an allele of BPH6. The identification of the resistance gene enriches the alleles of BPH6 and provides new materials for insect resistant rice breeding.

Key words: Oryza sativa, brown planthopper, BPH6, allele, physiological resistance mechanism

摘要：

【目的】发掘籼稻570011中抗褐飞虱主效基因，为培育抗虫水稻新品种提供基因资源。【方法】采用苗期集团法对抗性亲本570011和感虫亲本9311杂交后代F₃群体进行表型鉴定，结合F₂群体基因型，使用作图软件构建染色体的局部遗传连锁图，对目标区段抗性位点进行检测和遗传效应评估。采用实时荧光定量PCR (qRT-PCR)分析定位区间内最可能的候选基因，并对其基因组序列进行测序，比较对应CDS和氨基酸序列并进行系统进化树分析。【结果】籼稻570011在苗期对褐飞虱表现高抗，且对褐飞虱有明显的抗生性和趋避性。统计发现F₃群体抗虫株系数（抗性值<7）∶感虫株系数（抗性值≥7）为89∶35，卡方检验表明符合一对显性基因的分离规律。基因定位发现在第4染色体标记4M18.675和4M24.64之间的39 cM区域内检测到一个褐飞虱抗性位点，可能是已克隆基因BPH6的等位基因。qRT-PCR分析表明570011中Os04g35210（BPH6等位基因）在褐飞虱取食后表达量显著高于感虫材料9311。570011中Os04g35210基因与BPH6的CDS和氨基酸序列同源性分别达到99.08%和97.96%，系统进化树分析发现两者的亲缘关系最近，表明该抗性基因是BPH6的等位基因。【结论】籼稻材料570011是一份高抗褐飞虱种质，其抗性表型受一个主效基因控制。该基因的发掘丰富了BPH6的等位型和抗源来源，为抗虫水稻育种提供新抗源。

关键词: 水稻, 褐飞虱, BPH6, 等位基因, 生理抗性机制

CHENG Ling, HUANG Fugang, QIU Yipu, WANG Xinyi, SHU Wan, QIU Yongfu, LI Fahuo. Genetic Analysis and Identification of Brown Planthopper Resistance Gene in indica Rice Accession 570011[J]. Chinese Journal OF Rice Science, 2023, 37(3): 244-252.

程玲, 黄福钢, 邱一埔, 王心怡, 舒宛, 邱永福, 李发活. 籼稻材料570011抗褐飞虱基因的遗传分析及鉴定[J]. 中国水稻科学, 2023, 37(3): 244-252.

Figures/Tables 6

Fig. 1. Physiological resistance characterization of indica rice 570011 against BPH. A, Before BPH infestation; B, 9 d after BPH infestation; C, BPH host choice test; D, Number of survival BPH; E, BPH weight gain per BPH per 48 h; F, BPH honeydew weight per BPH per 48 h. * and ** indicate significant difference at the level of 5% or 1%, respectively.

Fig. 2. Distribution of BPH resistance scores of F3 mapping population derived from 9311 / 570011.

Fig. 3. qRT-PCR analysis of BPH6 allelic genes derived from indica rice 570011 and 9311. * indicate significant difference between two materials at the level of 5%.

Table 1. Resistance gene mapping with F3 population derived from 9311 / 570011.

标记 Marker	位置 Position/cM	LOD值 LOD score	PEV /%	A	D
4M18.675	0	1.77	3.0	0.44	−0.30
qBph4(t)	27	16.23	33.0	1.93	−1.64
4M24.64	39	2.26	4.0	1.20	−0.37
4M24.7	42	0.02	0.1	0.10	0.09
4M27.553	46	0.11	0.2	0.39	−0.20
4RM11491	47	0.84	1.5	0.90	−0.44

Fig. 4. Amino acid sequence comparison of Os04g35210 derived from indica rice 570011 and cloned gene BPH6.

Fig. 5. Phylogenetic tree developed with amino acid sequence of BPH6 and its homologous sequences. Box means amino acid sequences of cloned BPH6 and allelic gene derived from resistance accession 570011.

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