我国不同地区熊蜂短膜虫的感染情况及亲缘关系

doi:10.3864/j.issn.0578-1752.2019.06.014

Abstract

Abstract:

【Objective】The objective of this study is to clarify the prevalence of Crithidia bombi, an intestinal parasite of bumblebees, and its genetic relationship among different species in different regions.【Method】In this study, the infection status of 1 007 bumblebees from 25 species in four provinces (autonomous region) including Inner Mongolia, Gansu, Qinghai, and Sichuan was investigated, and the infection rate of C. bombi in different species and regions was analyzed by chi-square test (SPSS 22.0). PCR amplification, cloning and sequencing were carried out based on the ITS sequences of C. bombi. The specific ITS primers were used to amplify the total DNA in the intestines of bumblebees, and gel electrophoresis was performed. The infection status of bumblebee colony was determined by whether 675 bp of ITS gene fragment of C. bombi was amplified or not, and further sequencing was carried out to analyze the genetic relationship of C. bombi in different species and regions. 【Result】Out of all the samples investigated, 262 bumblebees of 20 species were infected by C. bombi, with an infection rate of 26.0%. Among them, Bombus festivus and B. pyrosoma had the highest infection rates, and B. sibiricus had the lowest infection rate. The infection rate of bumblebee was the highest in Qinghai Province and lowest in Inner Mongolia. The infection rate of male bee was significantly higher than that of worker bee and queen bee. The genetic relationship of most C. bombi was relatively close, except for B. melanurus and B. difficillimus in Gansu Province, and B. remotus, B. impetuosus, B. friseanus in Sichuan Province.【Conclusion】The infection of C. bombi is widespread in China, and the infection rate is different among different species, regions and castes of bumblebee. After a long-term of population evolution, the dominant species of bumblebee are relatively fixed in different regions, so the bumblebee species collected in different regions are different. The statistical analysis of the infection of C. bombi in different regions of China focused on the sample size ≥30, which are the dominant species in each region. Some bumblebees have not collected enough samples, but the infection rate is high according to the sample that we already get. There is host specificity in C. bombi, and C. bombi are adaptive to the infection of the host. In some regions of China, the genetic relationship between the C. bombi is relatively strong on the whole, but it is also affected by the geographical location and the species of bumblebee.

Key words: bumblebee (Bombus spp.), Crithidia bombi, internal transcribed space (ITS), infection rate, genetic relationship

TANG YuJie,WANG LiuHao,LI Kai,LI JiLian. Infection Status and Genetic Relationship of Crithidia bombi in Different Regions of China[J].Scientia Agricultura Sinica, 2019, 52(6): 1110-1118.

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Figures/Tables 7

Table 1

The species and number of bumblebees in different regions "

省（自治区） Province (autonomous region)	纬度 Latitude	经度 Longitude	海拔 Elevation (m)	蜂种 Species	采集数 Collection number	感染数 Infected number
内蒙古 Inner Mongolia	48°41.448′	122°45.290′	419	密林熊蜂B. patagiatus	41	5
	48°41.448′	122°45.290′	419	明亮熊蜂B. lucorum	4	3
	49°31.497′	117°16.951′	712	西伯熊蜂B. sibiricus	50	0
	49°28.953′	119°40.439′	602	盗熊蜂B. filchnerae	2	2
	49°28.953′	119°40.439′	602	藓状熊蜂B. muscorum	1	0
甘肃 Gansu	36°34.728′	102°58.077′	2096	黑尾熊蜂B. melanurus	7	1
	36°34.728′	102°58.077′	2096	西伯熊蜂B. sibiricus	1	1
	36°49.855′	102°39.003′	2210	火红熊蜂B. pyrosoma	19	13
	36°49.855′	102°39.003′	2210	明亮熊蜂B. lucorum	2	1
	36°49.855′	102°39.003′	2210	盗熊蜂B. filchnerae	3	1
	34°25.522′	102°17.784′	3454	稳纹熊蜂B. waltoni	7	0
	34°25.522′	102°17.784′	3454	拉熊蜂B. laesus	1	0
	34°42.456′	102°30.892′	3288	密林熊蜂B. patagiatus	52	12
	34°42.456′	102°30.892′	3288	兴熊蜂B. impetuosus	2	0
	38°09.402′	100°11.705′	2912	红束熊蜂B. rufofasciatus	103	15
	38°09.402′	100°11.705′	2912	苏氏熊蜂B. sushkini	3	2
	38°09.402′	100°11.705′	2912	伪猛熊蜂B. personatus	2	0
	38°09.402′	100°11.705′	2912	克什米尔熊蜂B. kashmirensis	2	1
	38°09.402′	100°11.705′	2912	越熊蜂B. supremus	3	1
	37°56.221′	100°57.951′	3418	猛熊蜂B. difficillimus	4	1
	37°59.971′	100°45.020′	3214	拉达克熊蜂B. ladakhensis	6	0
	37°59.971′	100°45.020′	3214	昆仑熊蜂B. keriensis	7	1
	38°13.043′	100°56.944′	2835	小雅熊蜂B. lepidus	3	0
青海 Qinghai	37°10.689′	102°03.091′	2596	明亮熊蜂B. lucorum	10	9
	37°10.689′	102°03.091′	2596	火红熊蜂B. pyrosoma	11	7
	37°10.689′	102°03.091′	2596	西氏熊蜂B. sichelii	5	4
	37°10.689′	102°03.091′	2596	小雅熊蜂B. lepidus	9	7
	37°10.689′	102°03.091′	2596	密林熊蜂B. patagiatus	1	0
	37°10.689′	102°03.091′	2596	兴熊蜂B. impetuosus	3	0
	37°41.408′	100°34.319′	3461	稳纹熊蜂B. waltoni	10	0
	37°41.408′	100°34.319′	3461	拉达克熊蜂B. ladakhensis	3	0
	37°41.408′	100°34.319′	3461	伪猛熊蜂B. personatus	2	0
	37°41.408′	100°34.319′	3461	红束熊蜂B. rufofasciatus	1	0
	37°41.408′	100°34.319′	3461	克什米尔熊蜂B. kashmirensis	1	0
四川 Sichuan	28°19.672′	103°07.999′	2062	兴熊蜂B. impetuosus	80	12
	28°19.672′	103°07.999′	2062	小雅熊蜂B. lepidus	12	2
	28°19.672′	103°07.999′	2062	三条熊蜂B. trifasciatus	8	5
	28°19.672′	103°07.999′	2062	白背熊蜂B. festivus	40	28
	28°19.672′	103°07.999′	2062	常熊蜂B. avanus	10	10
	30°02.905′	101°58.049′	2833	疏熊蜂B. remotus	159	32
	28°32.822′	102°10.638′	1785	弗里熊蜂B. friseanus	317	86
合计Total					1007	262

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References 28

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蜂种 Species	样本数（只） Sample number	感染率 Infection rate (%)	卡方值 χ²value	P值 P value
白背熊蜂B. festivus	40	70.0A	36.368	1.63E-09
火红熊蜂B. pyrosoma	30	66.7A	23.695	0.000001
弗里熊蜂B. friseanus	317	27.1B	0.057	0.812
疏熊蜂B. remotus	159	20.1BC	2.931	0.087
密林熊蜂B. patagiatus	94	18.1BC	3.178	0.075
红束熊蜂B. rufofasciatus	104	14.4C	7.274	0.007
兴熊蜂B. impetuosus	85	14.1C	6.323	0.012
西伯熊蜂B. sibiricus	51	2.0D	15.457	0.000084

蜂种 Species	四川Sichuan		甘肃Gansu		内蒙古Inner Mongolia
蜂种 Species	样本数（只） Sample number	感染率 Infection rate (%)	样本数（只） Sample number	感染率 Infection rate (%)	样本数（只） Sample number	感染率 Infection rate (%)
白背熊蜂B. festivus	40	70.0A	—	—	—	—
弗里熊蜂B. friseanus	317	27.1B	—	—	—	—
疏熊蜂B. remotus	159	20.1B	—	—	—	—
兴熊蜂B. impetuosus	80	15.0B	—	—	—	—
密林熊蜂B. patagiatus	—	—	52	23.1A	41	12.2A
红束熊蜂B. rufofasciatus	—	—	103	14.6A	—	—
西伯熊蜂B. sibiricus	—	—	—	—	50	0B

省（自治区）Province (autonomous region)	密林熊蜂B. patagiatus		火红熊蜂B. pyrosoma		稳纹熊蜂B. waltoni
省（自治区）Province (autonomous region)	样本数（只） Sample number	感染率 Infection rate (%)	样本数（只） Sample number	感染率 Infection rate (%)	样本数（只） Sample number	感染率 Infection rate (%)
内蒙古Inner Mongolia	41	12.2A	—	—	—	—
甘肃 Gansu	52	23.1A	19	68.4A	7	0A
青海 Qinghai	—	—	11	63.6A	10	0A

省（自治区）Province (autonomous region)	样本数（只） Sample number	感染率 Infection rate (%)	卡方值 χ²value	P值 P value
青海 Qinghai	56	48.2A	12.668	0.000372
四川 Sichuan	626	28.0B	0.457	0.499
甘肃 Gansu	227	22.0B	1.945	0.163
内蒙古Inner Mongolia	98	10.2C	12.655	0.000375

级型 Caste	样本数（只）Sample number	感染率 Infection rate (%)	卡方值 χ²value	P值 P value
雄蜂 Male	76	44.7A	12.467	0.000414
工蜂 Worker	929	24.5B	0.556	0.456
蜂王 Queen	2	0C	0.703	0.402

Infection Status and Genetic Relationship of Crithidia bombi in Different Regions of China

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[11]	WANG Kui-ling,MU Shao-hua,LIU Qing-chao,TANG Qi-he,LIU Qing-hua . Study on the Genetic Diversity of Some Camellia japonica (Naidong) Cultivars Based on AFLP Markers [J]. Scientia Agricultura Sinica, 2011, 44(3): 651-656 .
[12]	DING An-Ming, CUI Fa, LI Jun, ZHAO Chun-Hua, WANG Xiu-Qin, WANG Hong-Gang. QTL Analysis on Grain Yield per Plant and Plant Height in Wheat [J]. Scientia Agricultura Sinica, 2011, 44(14): 2857-2867 .
[13]	DUAN Yun-shang,JIANG Yan-hua,WANG Li-yuan,CHENG Hao,WANG Yu-hua,LI Xing-hui . Analysis of Genetic Diversity and Relationship of Tea Cultivars and Lines Suitable for Making Green and Black Tea Using SSR Markers [J]. Scientia Agricultura Sinica, 2011, 44(1): 99-109 .
[14]	JIAN Li,ZHU Li-quan . Analysis on Cymbidium kanran with SRAP Markers [J]. Scientia Agricultura Sinica, 2010, 43(15): 3184-3190 .
[15]	ZENG Li,ZHAO Liang-jun,SUN Jia,ZHAO Zi-gang,YANG-fan. Analysis of Genetic Relatedness of Genetic Resources of Tagetes as Revealed by ISSR [J]. Scientia Agricultura Sinica, 2010, 43(1): 215-222 .