【摘要】：瓜實蠅Bactrocera cucurbitae(Coquillett)是重要的農(nóng)業(yè)害蟲,生物防治實蠅技術—昆蟲不育技術[Sterile Insect Technique(SIT)]已越來越受到國內(nèi)外農(nóng)業(yè)專家的青睞,而SIT中的輻照過程削弱了靶標實蠅的體質(zhì)健康,關于這一方面的研究還鮮少有詳細的報道,本文研究了輻照對瓜實蠅遺傳區(qū)性品系[B.cucurbitae genetic sexing strain(GSS)]腸道菌的多樣性和相對豐度的影響,并分離、鑒定和篩選優(yōu)勢菌屬飼喂蟲源,在一定程度上降低了輻照的傷害。研究結果如下:1、以100Gy137Cs輻照瓜實蠅遺傳區(qū)性品系的5、8、11和13日齡的雌、雄成蟲,并采用基于16SrDNA的IlluminaMiSeq測序技術分別對其腸道菌群進行測序分析。結果表明從瓜實蠅雌、雄成蟲腸道中共檢測到4個菌門和19個主要菌屬,其中變形菌門Proteobacteria(占總量的50%～98%)、厚壁菌門Firmicutes(占總量的2%～47%)和普羅威登斯菌屬Providencia sp.、腸桿菌屬Enterobacter sp.分別是瓜實蠅GSS腸道中的優(yōu)勢菌門和菌屬;輻照顯著的減少了腸道中的微生物多樣性,降低了優(yōu)勢菌屬的相對豐度,這與瓜實蠅的性別和成蟲日齡無關。2、利用傳統(tǒng)的微生物分離技術從瓜實蠅雌、雄5日齡成蟲腸道中共分離得到7種菌屬,分別為:腸桿菌屬(37%),普羅威登斯菌屬(18.5%),克雷伯氏菌屬Klebsiella sp.(1.9%),芽孢桿菌屬Lysinibacillus sp.(1.9%),西地西菌屬 Cedecea sp.(24%),Bacterium sp.(11.1%)和未知屬名的其他菌屬other(5.6%)。3、利用腸桿菌屬作為瓜實蠅GSS幼蟲飼料的添加劑,用活菌(PI)、死菌(API)分別作為兩個處理組,用無菌(NPI)作為對照,飼喂寄主至化蛹后,分別測定瓜實蠅GSS的生物學和形態(tài)學指標,結果發(fā)現(xiàn)活菌組能顯著的增加瓜實蠅GSS的蛹重、頭寬和胸長,而死菌和活菌組都能顯著的提高雌、雄成蟲的耐壓存活率,它們提高的平均百分比分別是18%和4.72%(雌蟲),15%和11%(雄蟲)。因腸桿菌屬易分離和培養(yǎng),在所分離的菌屬中所占比例最大,而應用腸桿菌屬優(yōu)化幼蟲飼料有顯著的效果,結果證明腸桿菌屬是瓜實蠅GSS腸道中的優(yōu)勢菌屬,也是強化幼蟲飼料的益生菌屬。本研究成果將為未來開發(fā)利用營養(yǎng)菌作為瓜實蠅GSS幼蟲飼料的添加劑,從而在一定程度上修復輻照的損傷提供理論依據(jù),以期提高不育成蟲的質(zhì)量,為能高效的應用SIT技術打下基礎。
[Abstract]:Bactrocera cucurbitae (Coquillett) is an important agricultural pest. Biological control technology of fly, insect sterility technique [Sterile Insect Technique (SIT)] has been more and more favored by agricultural experts at home and abroad. However, the irradiation process in SIT weakens the physical health of the target fly, and there are few detailed reports on this aspect. The effects of irradiation on the diversity and relative abundance of intestinal bacteria in the genetic line [B.cucurbitae genetic sexing strain (GSS)] of melon fruit fly were studied, and the dominant bacteria were isolated, identified and screened, which reduced the damage caused by irradiation to some extent. The results were as follows: 1. 100Gy137Cs was used to irradiate the female, male and male adults of 5, 811 and 13 day old female and male adults of genetic region of fruit fly, and the intestinal flora was sequenced by IlluminaMiSeq sequencing technique based on 16SrDNA. The results showed that there were 4 phylum and 19 main genera in the intestinal tract of female and male adults of fruit fly, among which Proteobacteria was 50%. Firmicutes (2 / 47% of total) and Providencia sp., (Enterobacter sp.) It is the dominant phylum and genus in GSS intestinal tract of fruit fly, respectively. Irradiation significantly reduced the microbial diversity in the intestine and the relative abundance of the dominant genus, which had nothing to do with the sex of the fruit fly and the age of the adult. 2. The traditional microbiological isolation technique was used to isolate the female from the fruit fly. Seven species of bacteria were isolated from the intestinal tract of male adults at 5 days of age. They were Enterobacter (37%), Providence (18.5%), Klebsiella (1.9%) and Bacillus (Lysinibacillus sp. (1.9%). Cedecea sp. (24%), Bacterium sp. (11. 1%) and other unknown genus other (5. 6%). 3. Enterobacter was used as additive for GSS larvae of fruit fly and (PI), was used as a living bacterium. Dead bacteria (API) was used as two treatment groups, and sterile (NPI) was used as control. After feeding host to pupa, the biological and morphological indexes of GSS were measured respectively. The results showed that the live bacteria group could significantly increase the pupae weight of GSS. The head width and chest length were significantly increased in both dead and living bacteria groups, and the average percentage of increase was 18% and 4.72% (female), 15% and 11% (male), respectively. Because Enterobacterium is easy to isolate and culture, it accounts for the largest proportion in the isolated genus, and the application of Enterobacter to optimize larval feed has remarkable effect. The results show that Enterobacter is the dominant genus in the intestinal tract of GSS. It is also a probiotic genus that strengthens larval feed. The results of this study will provide a theoretical basis for the future development and utilization of nutritious bacteria as a feed additive for GSS larvae of fruit flies, thus repairing the damage caused by irradiation to a certain extent, in order to improve the quality of sterile adults. It lays the foundation for the efficient application of SIT technology.
【學位授予單位】：福建農(nóng)林大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S433

【參考文獻】

相關期刊論文 前10條

1 高帥;孫曉輝;;瓜實蠅的發(fā)生與綜合防治[J];蔬菜;2016年08期

2 駱米娟;張賀賀;陳家驊;杜迎剛;何龍艷;季清娥;;南亞實蠅成蟲腸道細菌的分離與鑒定[J];福建農(nóng)林大學學報(自然科學版);2016年01期

3 王洪秀;靳亮;陳慶隆;魏云輝;馬吉平;姚健;陳柳萌;鐘國祥;;橘小實蠅成蟲腸道可培養(yǎng)細菌群落結構分析[J];微生物學通報;2015年12期

4 程悅;左浩江;廖虹瑜;陳嘉熠;鄭田利;裴曉方;許欣;;腸道常見菌群熒光定量PCR檢測方法的建立[J];現(xiàn)代預防醫(yī)學;2014年23期

5 崔海輝;;蜜蜂消化道內(nèi)含物細菌群落組成的PCR-DGGE分析[J];浙江農(nóng)業(yè)科學;2014年05期

6 郭志強;農(nóng)軍;黃立飛;楊憲;;廣西絲瓜瓜實蠅的發(fā)生規(guī)律及綜合防治[J];安徽農(nóng)業(yè)科學;2014年02期

7 許波;楊云娟;李俊俊;唐湘華;慕躍林;黃遵錫;;宏基因組學在人和動物胃腸道微生物研究中的應用進展[J];生物工程學報;2013年12期

8 孫欣;高瑩;楊云鋒;;環(huán)境微生物的宏基因組學研究新進展[J];生物多樣性;2013年04期

9 劉莉揚;崔鴻飛;田埂;;高通量測序技術在宏基因組學中的應用[J];中國醫(yī)藥生物技術;2013年03期

10 王金濤;董永成;李宗鍇;黎少波;牛長纓;;昆蟲不育技術防治柑橘大實蠅研究概況[J];應用昆蟲學報;2013年03期

相關會議論文 前1條

1 季清娥;郭慶亮;陳家驊;;ME對橘小實蠅不育雄蟲交配能力的影響[A];第六屆核農(nóng)學青年科技工作者學術交流會暨中國核學會2011年學術年會核農(nóng)學分會論文集[C];2011年

相關博士學位論文 前2條

1 張南南;中國寡鬃實蠅亞科部分物種的形態(tài)分類及其分子系統(tǒng)發(fā)育研究[D];福建農(nóng)林大學;2012年

2 江昌木;瓜實蠅種群系統(tǒng)與檢疫防除綜合技術[D];福建農(nóng)林大學;2005年

相關碩士學位論文 前5條

1 崔守東;輻照導致橘小實蠅對性誘劑趨性變化機理的研究[D];福建農(nóng)林大學;2016年

2 蔡普默;橘小實蠅輻照卵作寄主繁殖阿里山潛蠅繭蜂的研究[D];福建農(nóng)林大學;2015年

3 吳玉華;瓜實蠅遺傳區(qū)性品系的基礎研究[D];福建農(nóng)林大學;2015年

4 楊焊;四種鱗翅目害蟲腸道細菌多樣性分析[D];南京農(nóng)業(yè)大學;2012年

5 張獻強;瓜實蠅的防治藥劑篩選及應用技術研究[D];廣西大學;2011年

，

本文編號：2405236