中国血吸虫病防治杂志
中國血吸蟲病防治雜誌
중국혈흡충병방치잡지
CHINESE JOURNAL OF SCHISTOSOMIASIS CONTROL
2014年
5期
526-530
,共5页
王海防%王怀位%程鹏%郭秀霞%杨培培%公茂庆
王海防%王懷位%程鵬%郭秀霞%楊培培%公茂慶
왕해방%왕부위%정붕%곽수하%양배배%공무경
中华按蚊%嗜血习性%rDNA-ITS2%遗传分化%标记-释放-重捕法
中華按蚊%嗜血習性%rDNA-ITS2%遺傳分化%標記-釋放-重捕法
중화안문%기혈습성%rDNA-ITS2%유전분화%표기-석방-중포법
Anopheles sinensis%Host-feeding preference%rDNA-ITS2%Genetic divergence%Mark-release-recapture method
目的：探讨同域分布而嗜血习性不同的中华按蚊种群是否存在基于rDNA-ITS2序列差异的遗传分化。方法在中华按蚊密度高的自然村和按蚊孳生地之间，同时设人饵和牛饵帐通宵诱捕大量野外按蚊种群；经过形态鉴定分离两组中华按蚊，并带回实验室常规饲养。选择一温湿度适宜的大型封闭温室作为雌蚊标记-释放-重捕技术的场所，将以上人饵组和牛饵组中华按蚊分别经红色和黄色荧光粉标记后在温室的中间一起释放，同时在温室两端分别设人饵帐和牛饵帐再次诱捕（重捕）中华按蚊。将两次均被人饵帐和牛饵帐诱捕的中华按蚊雌蚊分成嗜人血组和嗜牛血组，带回实验室饲养传代，并分别饲以其蚊虫人血和牛血。两组中华按蚊的子1代雌蚊再次使用上述标记-释放-重捕技术，筛选嗜人血和嗜牛血的中华按蚊品系；并对该两品系的中华按蚊rDNA-ITS2基因进行PCR扩增测序比对。结果野外亲代嗜人血组中华按蚊人饵帐和牛饵帐的重诱捕比例分别为54.07%（339/627）和45.93%（288/627），嗜牛血组牛饵帐和人饵帐的重捕比例分别为58.01%（409/705）和41.99%（296/705），两组亲代嗜血习性均趋向选择原吸血宿主（χ2=19.42，P<0.01）。子1代嗜人血组中华按蚊人饵帐和牛饵帐的重捕比例分别为63.43%（765/1206）和36.57%（441/1206），嗜牛血组牛饵帐和人饵帐的重捕比例分别为68.22%（1039/1523）和31.78%（484/1523），两组蚊虫群体嗜血习性有更显著的选择原吸血宿主的特性（χ2=271.69，P<0.01），表现出遗传分化现象。但PCR扩增测序比对结果显示，两品系蚊虫的rDNA-ITS2基因碱基序列无差异，均为469 bp。结论嗜血习性不同的中华按蚊同域种群未发生基于rDNA-ITS2序列的遗传分化。
目的：探討同域分佈而嗜血習性不同的中華按蚊種群是否存在基于rDNA-ITS2序列差異的遺傳分化。方法在中華按蚊密度高的自然村和按蚊孳生地之間，同時設人餌和牛餌帳通宵誘捕大量野外按蚊種群；經過形態鑒定分離兩組中華按蚊，併帶迴實驗室常規飼養。選擇一溫濕度適宜的大型封閉溫室作為雌蚊標記-釋放-重捕技術的場所，將以上人餌組和牛餌組中華按蚊分彆經紅色和黃色熒光粉標記後在溫室的中間一起釋放，同時在溫室兩耑分彆設人餌帳和牛餌帳再次誘捕（重捕）中華按蚊。將兩次均被人餌帳和牛餌帳誘捕的中華按蚊雌蚊分成嗜人血組和嗜牛血組，帶迴實驗室飼養傳代，併分彆飼以其蚊蟲人血和牛血。兩組中華按蚊的子1代雌蚊再次使用上述標記-釋放-重捕技術，篩選嗜人血和嗜牛血的中華按蚊品繫；併對該兩品繫的中華按蚊rDNA-ITS2基因進行PCR擴增測序比對。結果野外親代嗜人血組中華按蚊人餌帳和牛餌帳的重誘捕比例分彆為54.07%（339/627）和45.93%（288/627），嗜牛血組牛餌帳和人餌帳的重捕比例分彆為58.01%（409/705）和41.99%（296/705），兩組親代嗜血習性均趨嚮選擇原吸血宿主（χ2=19.42，P<0.01）。子1代嗜人血組中華按蚊人餌帳和牛餌帳的重捕比例分彆為63.43%（765/1206）和36.57%（441/1206），嗜牛血組牛餌帳和人餌帳的重捕比例分彆為68.22%（1039/1523）和31.78%（484/1523），兩組蚊蟲群體嗜血習性有更顯著的選擇原吸血宿主的特性（χ2=271.69，P<0.01），錶現齣遺傳分化現象。但PCR擴增測序比對結果顯示，兩品繫蚊蟲的rDNA-ITS2基因堿基序列無差異，均為469 bp。結論嗜血習性不同的中華按蚊同域種群未髮生基于rDNA-ITS2序列的遺傳分化。
목적：탐토동역분포이기혈습성불동적중화안문충군시부존재기우rDNA-ITS2서렬차이적유전분화。방법재중화안문밀도고적자연촌화안문자생지지간，동시설인이화우이장통소유포대량야외안문충군；경과형태감정분리량조중화안문，병대회실험실상규사양。선택일온습도괄의적대형봉폐온실작위자문표기-석방-중포기술적장소，장이상인이조화우이조중화안문분별경홍색화황색형광분표기후재온실적중간일기석방，동시재온실량단분별설인이장화우이장재차유포（중포）중화안문。장량차균피인이장화우이장유포적중화안문자문분성기인혈조화기우혈조，대회실험실사양전대，병분별사이기문충인혈화우혈。량조중화안문적자1대자문재차사용상술표기-석방-중포기술，사선기인혈화기우혈적중화안문품계；병대해량품계적중화안문rDNA-ITS2기인진행PCR확증측서비대。결과야외친대기인혈조중화안문인이장화우이장적중유포비례분별위54.07%（339/627）화45.93%（288/627），기우혈조우이장화인이장적중포비례분별위58.01%（409/705）화41.99%（296/705），량조친대기혈습성균추향선택원흡혈숙주（χ2=19.42，P<0.01）。자1대기인혈조중화안문인이장화우이장적중포비례분별위63.43%（765/1206）화36.57%（441/1206），기우혈조우이장화인이장적중포비례분별위68.22%（1039/1523）화31.78%（484/1523），량조문충군체기혈습성유경현저적선택원흡혈숙주적특성（χ2=271.69，P<0.01），표현출유전분화현상。단PCR확증측서비대결과현시，량품계문충적rDNA-ITS2기인감기서렬무차이，균위469 bp。결론기혈습성불동적중화안문동역충군미발생기우rDNA-ITS2서렬적유전분화。
Objective To investigate the existence of genetic divergence of sympatric populations of Anopheles sinensis of different feeding preferences based on the rDNA-ITS2 sequence differences. Methods A large number of wild anopheles popu-lations were trapped all night by man-baited net and calf-baited net that had been set up between high-density natural villages of An. sinensis populations and vector-breeding sites,from which two groups of An. sinensis were separated by morphological iden-tification and brought back to the lab for conventional breeding. A large closed greenhouse which temperature and humidity was appropriate was selected as research settings of mark-release-recapture methods by female mosquitoes ,in the center of which above An. sinensis populations baited by man and calf and respectively correspondingly marked by red and yellow phosphors were released in together,in each side of which An. sinensis were recaptured simultaneously by man-baited net and calf-baited net. An. sinensis populations trapped by man twice were brought back to the lab and bred with man-blood,correspondingly ones trapped by calf with calf-blood. Man-preferring and calf-preferring strains were screened respectively from An. sinensis which had been baited by man and calf by the mark-release-recapture methods after parent and F1 mosquitoes,and sequencing and aligning of both rDNA-ITS2 were conducted via PCR amplification. Results The recapture ratios of wild parental mosquitoes An. sinensis of man-preferring group by man-baited net and calf-baited net were 54.07%(339/627)and 45.93%(288/627)re-spectively,and ones of calf-preferring group by man-baited net and calf-baited net were 58.01%(409/705)and 41.99%(296/705)respectively. Two groups of parental mosquitoes trended towards selecting the original blood hosts in host-seeking prefer-ence(χ2=19.42,P<0.01). The recapture ratios of F1 mosquitoes An. sinensis of man-preferring group by man-baited net and calf-baited net were 63.43%(765/1 206)and 36.57%(441/1 206),and ones of calf-preferring group by man-baited net and calf-baited net were 68.22%(1 039/1 523)and 31.78%(484/1 523). Two groups of F1 mosquitoes had more significant characteris-tics of selecting the original blood hosts in host-seeking preference(χ2=271.69,P<0.01)and showed the genetic differentia-tion phenomenon,but the results of sequencing and aligning of the rDNA-ITS2 via PCR amplification showed no difference in base sequence between the two strains and both were 469 bp. Conclusions The genetic divergence based on the rDNA-ITS2 se-quence does not happen in An. sinensis sympatric populations of different feeding preferences.
