散白蚁卵巢发育及孤雌卵相关基因表达研究

发布时间：2018-05-29 08:49

本文选题：散白蚁 + 白蚁卵　；参考：《西北大学》2017年硕士论文

【摘要】：尖唇散白蚁Reticulitermes aculbialis和黄胸散白蚁R.flaviceps是我国重要的白蚁危害种类。本研究以尖唇散白蚁和黄胸散白蚁为材料,通过建立雌雌配对组(FF组)和雌雄配对组(FM组)人工巢群,采用显微组织观察、荧光染色、石蜡切片、荧光定量PCR、统计学分析等技术,系统地研究了两个种的产卵量、卵孔数、卵巢发育、未受精卵胚胎形成及发育相关基因的表达量,并对比了具有孤雌生殖能力的尖唇散白蚁和不具有孤雌生殖能力的黄胸散白蚁胚胎发育过程中在形态学与遗传学水平上的差异,旨在探究白蚁孤雌生殖的影响因子与调控机制。研究结论如下。(1)依据卵巢中最大卵母细胞的成熟程度,将卵巢分为五个阶段,A:前卵黄形成期卵母细胞阶段,B:早期卵黄形成期卵母细胞阶段,C:后期卵黄形成期卵母细胞阶段,D:成熟卵母细胞阶段,E:侧输卵管卵母细胞阶段。我们发现黄胸散白蚁FF组和FM组卵巢处在A和B阶段的总比率分别为53.33%和58.33%,尖唇散白蚁FF组和FM组处在C和D阶段的总比率分别为99.98%和75%,表明尖唇散白蚁卵巢发育与黄胸散白蚁卵巢相比更成熟。尖唇散白蚁和黄胸散白蚁FF组初建巢到产卵的时间间隔分别为35.12±2.59天和26.64±3.78天,八月份单个巢平均月产卵量分别为18.24±3.18,11.53±4.51,九月份单个巢平均月产卵量分别为11.16±4.26,3.67±1.24,表明尖唇散白蚁FF组产卵量要高于黄胸散白蚁FF组,且黄胸散白蚁九月份产卵量迅速降低。尖唇散白蚁FF组和FM组,黄胸散白蚁FF组和FM组的卵孔数分别为6.31±1.89,8.18±3.22,8.15±2.67,8.43±3.05,除黄胸散白蚁FF组卵孔数显著低于其他各组外,其他组间均无显著性差异。(2)尖唇散白蚁与黄胸散白蚁卵巢中成熟卵母细胞均存在一个卵核,刚产出孤雌卵存在2-5个卵核,表明两个种卵母细胞都在排卵过程中被激活且发生1-2次卵裂。尖唇散白蚁孤雌卵卵核数在产出5h为16-22个,10h为35-42个,24h为51-60个,48h为67-84个,卵裂速度逐渐减慢,且卵核向卵后端移动,3-5d形成胚盘,7-10天胚带形成,10-15d胚带延伸;黄胸散白蚁孤雌卵卵核数在产出5h为6-10个,48h为28-37个,5-7d为71-85个,且卵核移动至卵后端;黄胸散白蚁卵裂速度显著低于尖唇散白蚁,且8-10d时约86%的孤雌卵发育停滞在胚盘形成期,停滞卵卵黄细胞成块状聚集在卵的中央,卵两端呈中空状态。(3)我们测定了 8种与发育相关的基因在黄胸散白蚁FF组和尖唇散白蚁FF组中的表达量。对比同一个种内雌虫与卵巢的表达量,除尖唇散白蚁csf基因表达量在卵巢和成虫中无显著性差异外,黄胸散白蚁csf基因以及两个种中的cdc2,cyclin B,cyclin B3,mapkapk,cdc42,tnk2,ssp4基因在雌性生殖器官中的表达量都显著高于同种雌性成虫(P0.05),表明这些基因在卵巢中的表达上调;对比同一个种卵巢与产出的卵的基因表达量,两个种卵巢中cdc2,cyclin B,cyclin B3,mapkapk,cdc42,ssp4基因以及尖唇散白蚁中tnk2基因的表达量均显著高于其在相同种产出的卵中的表达量(P0.05),其余无明显差异,可能是由于卵产出后与发育相关的母源mRNA迅速降解。两个种间比较,尖唇散白蚁雌虫的cyclin,cyclinB3基因表达量显著高于黄胸散白蚁雌虫;cdc2,cyclin B,cyclin B3,mapkapk,cdc42,tnk2基因在尖唇散白蚁卵巢中的表达量显著高于黄胸散白蚁卵巢,与之相反csf基因在黄胸散白蚁卵巢中的表达量显著高于尖唇散白蚁卵巢,推测上述7个基因与卵巢发育和卵子成熟有关;尖唇散白蚁卵的tnk2基因含量显著高于黄胸散白蚁卵(P0.05),表明该基因可能与孤雌卵胚胎发育有关。
[Abstract]:Reticulitermes aculbialis and termite R.flaviceps are important species of termites in China. This study uses termite and yellow thorax termites as materials. By setting up female paired group (FF group) and male and female group (FM group) artificial nests, microtissue observation, fluorescent staining, paraffin section and fluorescence quantitative PCR are used. A systematic study of two species of oviposition, number of ovum, ovarian development, the formation of unfertilized eggs and the expression of genes related to development, and the comparison of the morphological and genetic water in the development of the embryo of the termites with the parthenogenetic ability of the parthenogenetic and the non parthenogenetic ability of the Yellow thorax termites. The conclusion is to explore the influencing factors and regulation mechanism of termite parthenogenesis. (1) according to the maturity of the largest oocyte in the ovary, the ovary is divided into five stages, the oocyte stage of A: before the yolk formation stage, the oocyte stage in the early yolk formation stage of B: and the oocyte stage in the late yolk formation stage of C:, D: mature oocyte stage and E: side oviduct oocyte stage. We found that the total ratio of group FF and FM in A and B stages were 53.33% and 58.33% respectively. The total ratio of the FF group and FM group at the C and D stages was 99.98% and 75% respectively. The time interval between the first built nest and the spawning in the FF group was 35.12 + 2.59 days and 26.64 + 3.78 days respectively. The average monthly egg production of the single nest in August was 18.24 + 3.18,11.53 4.51 respectively. The average monthly egg production of single nest in September was 11.16 + 4.26,3.67 + 1.24 respectively, indicating the oviposition amount of the FF group of termites apex. It was higher than group FF of termites yellow thorax, and the number of eggs of termite in September was reduced rapidly. The number of egg holes in group FF and FM, FF and FM in group FM of yellow thorax scattered termites were 6.31 + 1.89,8.18 + 2.67,8.43 + 2.67,8.43, respectively, and the number of ovum holes in group FF was significantly lower than that of the other groups. (2) There is an egg nucleus in the mature oocytes of the termite and the termite ovary of the Yellow thorax termite, and there are 2-5 egg nuclei in the parthenogenetic eggs, indicating that two ovules are activated in the process of ovulation and 1-2 cleavage. The number of egg nuclei of the oocyte of the apex termites is 16-22, 35-42, 51-60 24h, and 67-84 48h. The cleavage speed gradually slowed, and the egg nucleus moved to the back of the egg, 3-5d formed the embryo disc, formed the embryo zone in 7-10 days and extended the 10-15d embryo zone; the number of egg nuclei of the oocyte of termites of termites yellow thorax was 6-10, 28-37 48h and 71-85 5-7d, and the egg nucleus moved to the back end of the egg; the cleavage rate of the Yellow thorax scattered white ants was significantly lower than that of the apex labial termites, and about 86% 8-10d when 8-10d. The development of parthenogenetic eggs stagnated at the stage of embryonic disc formation, and the stagnant oval cells were lumped in the center of the egg, and the two ends of the eggs were hollow. (3) we measured the expression of 8 developmental related genes in the group FF of termites yellow thorax and the FF termites of the apex labial termites. The expression of the female and ovary in the same species was compared, and the CSF base of the termites was compared to the apex labial termites. The expression of CSF gene in yellow thorax termites and cdc2, cyclin B, cyclin B3, mapkapk, Cdc42, tnk2, Ssp4 genes in female reproductive organs were significantly higher than those of the same female adult (P0.05) in the two species, and the expression of these genes in the ovary was up. The expression of CDC2, cyclin B, cyclin B3, mapkapk, Cdc42, Ssp4, and sharp labial termites were significantly higher in the expression of tnk2 genes in the two species of ovaries than in the eggs produced in the same species (P0.05), and the rest was not significantly different. The expression of cyclin and cyclinB3 gene in the cyclin and cyclinB3 gene of termites was significantly higher than that of termites of yellow thorax; cdc2, cyclin B, cyclin B3, mapkapk, Cdc42, tnk2 genes were significantly higher in the ovaries of the termite ovaries than the Yellow thorax termite ovary, and the expression of the reverse CSF gene in the Yellow thorax termite ovary was significant. The above 7 genes are related to ovarian development and egg maturation, and the tnk2 gene content of termite eggs is significantly higher than that of yellow termites (P0.05), indicating that the gene may be related to the development of parthenogenetic eggs.
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q963

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