Analysis of Genetic Diversity of Nosema bombycis in Yunnan Sericulture Region

doi:10.19586/j.2095-2341.2024.0019

Current Biotechnology ›› 2024, Vol. 14 ›› Issue (4): 631-639.DOI: 10.19586/j.2095-2341.2024.0019

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Analysis of Genetic Diversity of Nosema bombycis in Yunnan Sericulture Region

Yonghong ZHANG¹(), Zhenguo SU¹, Jiafu LUO¹, Chunfeng LI²(), Baolin AO³

^1.Institute of Sericulture and Apiculture，Yunnan Academy of Agricultural Science，Yunnan Mengzi 661101，China
^2.Chongqing Key Laboratory of Microsporidia Infection and Control，Southwest University，Chongqing 400715，China
^3.Mulberry-silkworm Service Center of Luliang County，Yunnan Luliang 655600，China

Received:2024-02-04 Accepted:2024-04-24 Online:2024-07-25 Published:2024-08-07
Contact: Chunfeng LI

云南蚕区家蚕微孢子虫遗传多样性分析

张永红¹(), 苏振国¹, 罗家福¹, 李春峰²(), 敖宝林³

^1.云南省农业科学院蚕桑蜜蜂研究所，云南蒙自 661101
^2.西南大学，微孢子虫感染与防控重庆市重点实验室，重庆 400715
^3.陆良县蚕桑服务中心，云南陆良 655600

通讯作者: 李春峰
作者简介:张永红 E-mail: zhang200503@126.com；
基金资助:
云南省农业科学院预研项目(2024KYZX-01);重庆市重点实验室开放课题项目(CKMIC202102)

Abstract

Abstract:

Nosema bombycis， a crucial pathogen causing silkworm pebrin， poses a significant threat to sericulture production. This study aimed to investigate the genetic diversity within N. bombycis species， providing valuable insights for the prevention and control of silkworm pebrin in the Yunnan sericultural region. We collected infected silkworm samples from diverse rearing areas in Yunnan Province， isolated and purified the Bombyx mori larvae， extracted their genomes， cloned the small subunit ribosomal DNA （SSU rDNA） and internal transcribed spacer （ITS） sequences， and conducted comprehensive bioinformatics analyses. Results revealed that the SSU rDNA sequences of N. bombycis isolated in the Yunnan silkworm region exhibit over 99% homology， with a genetic distance of less than 0.006. However， these isolates displayed variations in length and multiple loci， indicating varying degrees of polymorphism. In contrast， the rDNA-ITS sequences exhibited significant genetic differences， characterized by multiple base insertions or deletions， as well as single base substitutions and inversions. Utilizing the SSU rDNA and ITS sequences， we constructed a phylogenetic tree， which revealed genetic differentiation among the isolated strains of N. bombycis in the Yunnan sericultural region. Notably， the results indicated no direct correlation between the genetic relationships of these populations and their geographical locations. This study has significantly enriched our understanding of the intra-species genetic diversity of N. bombycis in the Yunnan silkworm-raising region， providing a valuable foundation for further research on silkworm pebrin prevention and control strategies.

Key words: Nosema bombycis, genetic diversity, ribosomal small subunit, transcribed spacer

摘要：

家蚕微孢子虫（Nosema bombycis）是蚕业生产上一种重要病害——微粒子病的病原体。探讨家蚕微孢子虫种内的遗传多样性，可为云南蚕区家蚕微粒子病的防控提供参考依据。从云南省不同养蚕地区收集了感染微孢子虫的病蚕样品，分离纯化家蚕微孢子虫并提取基因组，克隆SSU rDNA（small subunit ribosomal DNA）和ITS（internal transcribed spacer）序列并进行生物信息学分析。结果发现，云南蚕区Nosema bombycis 分离株SSU rDNA序列同源性高达99%以上，遗传距离小于0.006，它们在长度和多个位点存在差异，呈现不同程度的多态性；ITS遗传差异较为显著，序列中存在多碱基的插入或缺失、单碱基的转换和颠换。基于SSU rDNA和rDNA-ITS序列构建系统发生树，结果显示，云南蚕区家蚕微孢子虫分离株系间存在遗传分化，种群间亲缘关系与地理位置无直接联系。研究结果丰富了云南蚕区家蚕微孢子虫的种内遗传多样性。

关键词: 家蚕微孢子虫, 遗传多样性, 核糖体小亚基, 转录间隔区

CLC Number:

Yonghong ZHANG, Zhenguo SU, Jiafu LUO, Chunfeng LI, Baolin AO. Analysis of Genetic Diversity of Nosema bombycis in Yunnan Sericulture Region[J]. Current Biotechnology, 2024, 14(4): 631-639.

张永红, 苏振国, 罗家福, 李春峰, 敖宝林. 云南蚕区家蚕微孢子虫遗传多样性分析[J]. 生物技术进展, 2024, 14(4): 631-639.

Figures/Tables 9

References 31

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名称	宿主	SSU rDNA GenBank登录号	ITS GenBank登录号	地理来源
YNMZCB1	Bombyx mori	OQ970549	OQ974967	云南省蒙自市
YNMZCB2		OQ970550	OQ974968	云南省蒙自市
YNMZLQ		OQ970551	OQ974969	云南省蒙自市
YNCXDY		OR597059	OR597057	云南省大姚县
YNQJLL		OR597056	OR597058	云南省陆良县
YN1		JF334596	JF443647	云南省
YN2		JF334597	JF443648	云南省
YN3		JF334598	JF443649	云南省
YN4		JF334599	JF443650	云南省
SC1		JF334592	JF443631	四川省
CQ1		EU350391	EU350394	重庆市
GX1		JF443547	JF443600	广西壮族自治区
GD1		JF334582	JF443619	广东省
AH		GQ334399	—	安徽省
JS		AY616662	—	江苏省
TW		AY209011	AY209011	台湾省
J-IK		AY259631	AY259631	日本茨城县
Nosema ceranae	Apis mellifera	DQ486027	—	台湾省
Nosema apis	Apis cerana	U97150	—	美国加利福尼亚州
Nosema sp.SC	Samia cynthia ricini	FJ767862	—	江苏省
Nosema sp.PX1	Plutella xylostae	AY960986	—	台湾省
Nosema sp.C01	Pieris rapae	AY383655	—	江苏省
Nosema plutellae	Plutella xylostea	AY960987	—	台湾省
Nosema spodoptrae	Spodoptera litura	AY747307	—	台湾省
Beauveria bassiana	Blattaria	EU334679	—	加拿大魁北克省
Nosema antheraeae	Antherea pernyi	—	DQ073396	河南省
Nosema disstriae	Malacasoma disstria	—	EU221229	加拿大安大略省

名称	宿主	SSU rDNA GenBank登录号	ITS GenBank登录号	地理来源
YNMZCB1	Bombyx mori	OQ970549	OQ974967	云南省蒙自市
YNMZCB2		OQ970550	OQ974968	云南省蒙自市
YNMZLQ		OQ970551	OQ974969	云南省蒙自市
YNCXDY		OR597059	OR597057	云南省大姚县
YNQJLL		OR597056	OR597058	云南省陆良县
YN1		JF334596	JF443647	云南省
YN2		JF334597	JF443648	云南省
YN3		JF334598	JF443649	云南省
YN4		JF334599	JF443650	云南省
SC1		JF334592	JF443631	四川省
CQ1		EU350391	EU350394	重庆市
GX1		JF443547	JF443600	广西壮族自治区
GD1		JF334582	JF443619	广东省
AH		GQ334399	—	安徽省
JS		AY616662	—	江苏省
TW		AY209011	AY209011	台湾省
J-IK		AY259631	AY259631	日本茨城县
Nosema ceranae	Apis mellifera	DQ486027	—	台湾省
Nosema apis	Apis cerana	U97150	—	美国加利福尼亚州
Nosema sp.SC	Samia cynthia ricini	FJ767862	—	江苏省
Nosema sp.PX1	Plutella xylostae	AY960986	—	台湾省
Nosema sp.C01	Pieris rapae	AY383655	—	江苏省
Nosema plutellae	Plutella xylostea	AY960987	—	台湾省
Nosema spodoptrae	Spodoptera litura	AY747307	—	台湾省
Beauveria bassiana	Blattaria	EU334679	—	加拿大魁北克省
Nosema antheraeae	Antherea pernyi	—	DQ073396	河南省
Nosema disstriae	Malacasoma disstria	—	EU221229	加拿大安大略省

分离株	核苷酸位点
分离株	79	102	294	445	471	558	565	591	640	651	696	703	823	844	861
YNMZCB1	G	G	T	G	—	T	G	G	—	C	A	T	G	A	C
YNMZCB2	G	G	C	G	T	T	A	G	—	T	A	C	A	G	T
YNMZLQ	A	G	C	G	—	T	A	G	—	T	A	C	G	G	C
YNCXDY	G	G	C	C	—	T	A	G	—	T	A	C	G	G	C
YNQJLL	G	G	C	G	—	T	A	G	—	T	A	C	G	G	C
JS	G	A	C	G	—	G	A	A	C	C	T	C	G	G	C

分离株	核苷酸位点
分离株	79	102	294	445	471	558	565	591	640	651	696	703	823	844	861
YNMZCB1	G	G	T	G	—	T	G	G	—	C	A	T	G	A	C
YNMZCB2	G	G	C	G	T	T	A	G	—	T	A	C	A	G	T
YNMZLQ	A	G	C	G	—	T	A	G	—	T	A	C	G	G	C
YNCXDY	G	G	C	C	—	T	A	G	—	T	A	C	G	G	C
YNQJLL	G	G	C	G	—	T	A	G	—	T	A	C	G	G	C
JS	G	A	C	G	—	G	A	A	C	C	T	C	G	G	C

	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	名称
1		0.006	0.005	0.005	0.004	0.010	0.011	0.006	0.009	0.006	0.004	0.006	0.011	0.006	0.006	0.006	0.004	YNMZCB1
2	99.4		0.002	0.002	0.002	0.006	0.006	0.000	0.006	0.004	0.003	0.002	0.008	0.004	0.006	0.002	0.002	YNMZCB2
3	99.5	99.8		0.002	0.001	0.007	0.007	0.002	0.006	0.003	0.001	0.002	0.007	0.003	0.005	0.002	0.001	YNMZLQ
4	99.5	99.8	99.8		0.001	0.007	0.007	0.002	0.006	0.003	0.002	0.002	0.007	0.003	0.005	0.002	0.001	YNCXDY
5	99.6	99.8	99.9	99.9		0.006	0.006	0.002	0.005	0.002	0.002	0.002	0.006	0.002	0.004	0.002	0.000	YNQJLL
6	99.0	99.4	99.3	99.3	99.4		0.011	0.006	0.011	0.008	0.007	0.006	0.012	0.008	0.010	0.006	0.006	YN1
7	98.9	99.4	99.3	99.3	99.4	98.9		0.006	0.011	0.009	0.008	0.006	0.013	0.006	0.011	0.006	0.006	YN2
8	99.4	100.0	99.8	99.8	99.8	99.4	99.4		0.006	0.004	0.003	0.002	0.008	0.004	0.006	0.002	0.002	YN3
9	99.1	99.4	99.4	99.4	99.5	98.9	98.9	99.4		0.007	0.006	0.006	0.011	0.007	0.009	0.006	0.005	YN4
10	99.4	99.6	99.7	99.7	99.8	99.2	99.1	99.6	99.3		0.004	0.004	0.007	0.005	0.006	0.004	0.002	SC1
11	99.6	99.7	99.9	99.8	99.8	99.3	99.2	99.7	99.4	99.6		0.003	0.008	0.004	0.006	0.003	0.002	CQ1
12	99.4	99.8	99.8	99.8	99.8	99.4	99.4	99.8	99.4	99.6	99.7		0.008	0.004	0.006	0.002	0.002	GX1
13	98.9	99.2	99.3	99.3	99.4	98.8	98.7	99.2	98.9	99.3	99.2	99.2		0.009	0.009	0.008	0.006	GD1
14	99.4	99.6	99.7	99.7	99.8	99.2	99.4	99.6	99.3	99.5	99.6	99.6	99.1		0.006	0.004	0.002	AH
15	99.4	99.4	99.5	99.5	99.6	99.0	98.9	99.4	99.1	99.4	99.4	99.4	99.1	99.4		0.006	0.004	JS
16	99.4	99.8	99.8	99.8	99.8	99.4	99.4	99.8	99.4	99.6	99.7	99.8	99.2	99.6	99.4		0.002	TW
17	99.6	99.8	99.9	99.9	100.0	99.4	99.4	99.8	99.5	99.8	99.8	99.8	99.4	99.8	99.6	99.8		J-IK

Analysis of Genetic Diversity of Nosema bombycis in Yunnan Sericulture Region

云南蚕区家蚕微孢子虫遗传多样性分析

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Figures/Tables 9

References 31

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分离株	长度/bp	GC含量/%
YNMZCB1	187	15.51
YNMZCB2	186	19.35
YNMZLQ	180	20.56
YNCXDY	184	21.20
YNQJLL	181	18.23
YN1~YN18	179~192	15.51~21.11
SC1~SC16	181~186	16.67~18.68
GX1~GX19	179~190	15.51~21.05
GD1~GD12	179~190	17.13~20.54
CQ1~CQ10	180~187	15.51~20.54

[1]	Xiaorui YAN, Guozhen XUE, Xiaoxia YANG, Yu WANG, Chenhui DU, Shuosheng ZHANG, Jiquan LIU. ISSR Genetic Diversity Analysis of Bupleurum chinense from Different Populations [J]. Current Biotechnology, 2023, 13(6): 919-924.
[2]	Qinxue GAO, Wenke YANG, Yonggang MENG, Nashier YASHENGJIANG, Kuwaxi MAIMAITITUERGAN. Genetic Diversity of Xinjiang Kizilsu Kirghiz Region Pamir Yaks Based on Mitochondrial COⅠ Gene [J]. Current Biotechnology, 2022, 12(4): 568-576.
[3]	LI Jing1, WANG Li1, DAO Min2, HU Ping1, XIAO Yi1, HAN Jianlin1, WANG Yutao1*. Genetic Diversity and Phylogenetic Analysis on mtDNA Cytb in Yak of Karakorum-Pamir Region [J]. Curr. Biotech., 2019, 9(5): 509-517.
[4]	ZHANG Zhi-ming1, TANG Cai-guo2,3, YANG San-wei1, QIAO Lin-yi1, CHANG Jian-zhong1, ZHAO Cui-rong4,5, ZHENG Jun1. Advances on Gene Discovery and Utilization of Wild Relatives of Triticum aestivum L. [J]. Curr. Biotech., 2016, 6(5): 305-311.
[5]	DONG Zhi-gang1§, LIU Zheng-hai2§, LI Xiao-mei1, . Application of SSR Markers in Variety Identification and Genetic Breeding of Vitis vinifera [J]. Curr. Biotech., 2016, 6(2): 137-140.
[6]	HE Zhen1, YUN Xiao-dong1, WU Kai1, JI Hu-tai2, CHANG Jian-zhong1, QIAO Lin-yi1, ZHENG Jun2. Progress on Germplasm Resources Genetic Diversity of Soybean [J]. Curr. Biotech., 2015, 5(2): 103-108.
[7]	WANG Shi-feng, LI Jing, WANG Yu-tao. Genetic Diversity and Phylogeny Analysis of Duolang Sheep Based on Mitochondrial DNA-loop Variation [J]. Curr. Biotech., 2014, 4(6): 429-434.
[8]	HE Hu-yi, TAN Guan-ning, HE Xin-min*, HE Hai-wang, LI Li-shu, TANG Zhou-ping, WANG Hui. Application of Several PCR-based Molecular Markers in Sweet Potato Genetic Diversity Researches [J]. Curr. Biotech., 2014, 4(4): 245-250.
[9]	WANG Han-yu1, Du Yan-wei2, LI Ping2, ZHANG Xi-wen2, ZHU Jing-ying1, ZHAO Jin-feng2, YU Ai-li1*. Genetic Relationship of 40 Maize Inbred Lines Based on ISSR [J]. journal1, 2011, 1(3): 214-218.