赤拟谷盗和杂拟谷盗的生殖隔离研究

赤拟谷盗和杂拟谷盗是两个同域发生的近缘储粮害虫。为明确它们的生殖隔离程度和机制,本研究比较分析了赤拟谷盗和杂拟谷盗雄虫对同种和异种雌虫的交配选择;赤拟谷盗与杂拟谷盗交配后,测定了精子在异种雌体内的存活情况;将赤拟谷盗和杂拟谷盗进行正反杂交,研究其F1代、F2代和回交代杂种是否产生及其雌雄比。结果显示,赤拟谷盗和杂拟谷盗雄虫对同种雌虫的爬跨率和交配率高于其对异种雌虫的爬跨率和交配率,表明赤拟谷盗和杂拟谷盗种间性隔离不完全;赤拟谷盗和杂拟谷盗交配后精子在异种雌体内是存活的,杂交所产的F1代卵为受精卵,说明交配后完成了精子传送和受精过程,表明种间机械隔离和配子隔离不完全;赤拟谷盗和杂拟谷盗的杂种F1代自交和回交产生了F2代和回交代,表明种间不存在杂种不育;一些杂交组合产生的F2代和回交代数量少且存在雌雄性比偏离,表明种间存在部分杂种不活和杂种衰败。本研究明确了赤拟谷盗和杂拟谷盗的生殖隔离机制,这对于揭示它们的物种进化关系有着重要意义。     

赤拟谷盗微卫星多态性的研究

为了研究赤拟谷盗种群的遗传结构,采用微卫星分子标记技术,对4个不同地理种群共120个个体的遗传多样性水平及种群遗传结构进行了研究.通过提取基因组DNA,用5对微卫星引物进行PCR扩增、电泳分析、凝胶成像系统分析,结果表明CB337218、DN648427、DN647734、EB754173、EB750288这5个位点的等位基因数分别为4、4、3、6和3, 4个不同地理种群赤拟谷盗的遗传杂合度平均值为0.4379,4个不同地理种群赤拟谷盗间的多态信息含量各在0.3771～0.5089之间,都达到中度多态性或高度多态性水平.为进一步研究赤拟谷盗的成灾机理提供了分子遗传学的基础.     

特定电磁波(TDP)对赤拟谷盗繁殖性能的影响

TDP辐射器经通电,产生一条2～50μm电磁波谱。该波自问世10多年来,在人和动物的疾病防治及其他生物学效应方面,均有实用价值。TDP辐射对家畜繁殖功能有促进和调节作用,前已有报道［1,2］。对昆虫赤拟谷盗TriboliumcastaneumHerbst的影响本实验尚属首次,赤拟谷盗,因其世代短,繁殖力强,性染色体雌虫为XX雄虫为XY,踊期可鉴别雌雄,故在遗传学研究中常作“导航试验昆虫”[3]。本试验目的在于观察TDP对赤拟谷盗繁殖功能的影响。1实验方法选野生型SH系中赤拟谷盗10个家系的蛹,鉴别雌雄,分别放入培养箱中（温度32℃,相对湿度60…     

温度对赤拟谷盗爬行和起飞活动的影响

在不同温度下研究了赤拟谷盗Tribolium castaneum(Herbst)在固体物面、设定粮面、温度均匀的粮(小麦)柱和具有温度梯度粮柱内爬行扩散速度、起飞温度等.主要结果为:赤拟谷盗在17℃开始有爬行为,25℃开始有飞行行为,在15～25℃时水平爬行速度和15 ～ 20℃时竖直运动速度与温度呈显著正相关,在无障...     

赤拟谷盗模拟种公畜选择──常规与改进方法的比较

本实验用实验昆虫赤拟谷盗两个世代各３０个家系的８５９和７２６只蛹的２３日龄蛹长和蛹宽作为选择性状模拟种公畜选择,对常规方法和改进方法进行比较。以Ｈｅｎｄｅｒｓｏｎ方法Ⅲ和综合指数作为常规方法,用约束最大似然法（ＲＥＭＬ）和ＢＬＵＰ估计方差组分和育种值为改进方法。结果表明,常规法估计遗传参数在两世代中不稳定,而改进方法弥补此缺点。对两世代个体和家系育种值排队,结果表明两种方法在前几名无差别,说明在选择强度大（留种率低）时,两种方法效果基本一致。     

赤拟谷盗全基因组和EST中微卫星的丰度

微卫星是近年大力开发的一种分子标记,为了推进赤拟谷盗Tribolium castaneum(Herbst)遗传学相关研究,对赤拟谷盗全基因组和EST中由1~6个碱基重复单元组成的简单序列重复进行分析,进而对其微卫星的丰度和分布进行比较分析。微卫星在赤拟谷盗EST中的分布频率为1/0.87kb,其中单碱基重复序列占71.25%,是最丰富的重复单元,而六、三、四、二,五碱基重复单元序列分别占23.93%,2.94%,1.56%,0.17%,0.15%。全基因组中微卫星的分布频率为1/3.65kb,其中六碱基重复序列占61.96%,是最丰富的重复单元,而三,四,一,五,二碱基重复单元序列分别占14.35%,13.75%,4.68%,3.60%,1.69%。同时发现富含A和T碱基的微卫星占主导地位,富含G和C碱基的微卫星数量较少。进一步的分析显示,微卫星在每条染色体上的丰度存在很大的相似性。     

温湿度对赤拟谷盗成虫飞行能力的影响

本文利用昆虫飞行磨研究了温度和湿度变化对赤拟谷盗Tribolium castaneum(Herbst)雌、雄成虫飞行能力的影响;分别测定了23℃、26℃、29℃、32℃和35℃等5个温度,45%、65%和85%等3个湿度条件下,赤拟谷盗雌、雄成虫的飞行距离、时间和速度等反应昆虫飞行能力的参数。单次飞行距离、单次飞行时间、单次飞行速度、24 h累计飞行距离及时间分析结果表明,环境温度和湿度对赤拟谷盗雌、雄成虫的飞行能力有不同程度的影响,环境温湿度过高或过低均不利于赤拟谷盗雌、雄成虫飞行;温度为32℃时,赤拟谷盗雌成虫的单次飞行距离、飞行时间、飞行速度、24 h累计飞行距离和飞行时间分别为0.925 m、9.616 s、0.023 m/s、0.979 km和2.886 h;雄成虫的单次飞行距离、飞行时间、飞行速度、24 h累计飞行距离和飞行时间分别为0.638 m、6.035 s、0.014 m/s、0.674 km和3.175 h。在相对湿度45%、65%和85%时,雌成虫的单次飞行时间分别为6.998 s、9.616 s和6.431 s;雄成虫的单次飞行时间分别为3.163 s、6.035 s和0.208 s。飞行速度与相对湿度的关系与飞行时间相似。从飞行能力参数来看,雌虫的飞行能力优于雄虫。     

冰核真菌削弱赤拟谷盗抗寒力的初步研究

赤拟谷盗Tribolium castaneum是不耐结冰的害虫,在冬季它通过降低过冷却点以避免结冰造成的致命伤害。冰核活性细菌能显著提高昆虫的过冷却点,使之在较高的零下温度发生结冰。试验证明冰核活性真菌也能显著提高赤拟谷盗的过冷却点。对照组平均过冷却点为-14.9℃。用10 g/L的冰核真菌制剂喷洒虫体,风干后测定,平均过冷却点提高到-4.8℃。用0.1 g/L处理后至少在7天内过冷却点保持较高。这些结果表明冰核真菌可能成为一种在冬季使用的、防治不耐结冰害虫的促冻杀虫剂。     

磷化氢对赤拟谷盗成虫体内CAT和SOD活性的影响

用赤拟谷盗Tribolium castaneum(Herbst)成虫,研究PH3对其体内超氧化物岐化酶(SOD)和过氧化氢酶(CAT)活性的影响。结果表明PH3熏蒸后,4种不同地理种群的成虫体内,CAT酶活性都有所降低。种群NGD1降低最多,接近57%,而NGD4则只降低了19%,NGD2和NGD3的降低都在40%左右。不同地理种群的成虫体内,SOD酶活性在熏蒸后都有所增加,种群NGD1增加幅度最大,NGD2增加幅度最小。SOD和CAT活性的变化幅度与成虫对PH3产生的抗性有关,抗性越大,酶活性变化越大。     

不同地理种群赤拟谷盗肠道细菌群落多样性分析

【目的】了解储粮害虫赤拟谷盗Tribolium castaneum (Herbst)成虫肠道细菌群落组成及多样性。【方法】利用Illumina MiSeq高通量测序技术对3个地理种群的赤拟谷盗雄虫肠道细菌16S rDNA的V3-V4变异区进行测序分析。【结果】从赤拟谷盗肠道样本中共测得465 293条reads,聚类为113个OTU,共注释到8门、15纲、33目、52科和79属的细菌。其中,优势菌门包括变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和放线菌门(Actinobacteria)等,优势菌属包括假单胞菌属(Pseudomonas)、肠杆菌科未鉴定属(Unclassified-f-Enterobacteriaceae)、不动杆菌属(Acinetobacter)、肠球菌属(Enterococcus)和根瘤菌属(Rhizobium)等。3个地理种群的赤拟谷盗肠道细菌的Shannon指数、Simpson指数、Ace指数和Chao指数分别为1.5-1.7、0.3、79.7-133.2和76.4-85.2。【结论】不同地理种群的赤拟谷盗成虫的肠道细菌群落组成存在显著差异。     

Identification of microsatellite markers in the red flour beetle,Tribolium castaneum

We isolated and characterized microsatellite markers for the red flour beetle Tribolium castaneum, an important model species for studies in various areas of evolutionary biology and ecology. A microsatellite‐enriched genomic library was constructed and screened with single stranded oligonucleotide probes [(CCT)₁₇, (AAT)₁₇ and (CAG)₁₇]. Forty‐five primer pairs were designed of which 19 pairs produced successful amplification. Polymorphism screening involved beetles from five beetle strains and revealed 15 polymorphic and four monomorphic markers. The development of polymorphic microsatellite markers will facilitate future ecological and genetic studies involving T. castaneum beetles.     

The effects of Farinocystis tribolii on the growth and development of the flour beetle Tribolium castaneum

Farinocystis tribolii multiplied only in the fatbody of Tribolium castaneum larvae. In the advanced stages of infection, the fatbody was destroyed and the albuminoid granules were completely depleted. Larvae in late stages of infection were “C” shaped. Diseased pupae were flat and straight with the head and mouth parts not completely flexed with the pupal body. Infected pupae failed to develop into adults. Heavily infected adults were distended. Larvae of third, fourth, and fifth instars, although succumbing to the infection ultimately, continued to live even after the healthy larvae of the same age had entered pupation, indicating a prolongation of larval period. Molting was either affected or delayed in infected insects. F. tribolii infection induced a juvenile hormonal effect producing larval-pupal and pupal-adult intermediate forms. Ether extract of spores of F. tribolii when applied on pupae of T. castaneum and final instar nymphs of Dysdercus cingulatus produced adultoids.     

Characterization and functional analysis of hsp18.3 gene in the red flour beetle,Tribolium castaneum

Small heat shock proteins (sHSPs) are diverse and mainly function as molecular chaperones to protect organisms and cells from various stresses. In this study, hsp 18.3, one Tribolium castaneum species-specific shsp、has been identified. Quantitative real-time polymerase chain reaction illustrated that Tchsp 18.3 is expressed in all developmental stages, and is highly expressed at early pupal and late adult stages, while it is highly expressed in ovary and fat body at the adult period. Moreover, it was up-regulated 4532 ± 396-fold in response to enhanced heat stress but not to cold stress;meanwhile the lifespan of adults in ds-Tchsp 18.3 group reduced by 15.8% from control group under starvation. Laval RNA interference (RNAi) of Tchsp 18.3 caused 86.1%±4.5% arrested pupal eclosion and revealed that Tchsp 18.3 played an important role in insect development. In addition, parental RNAi of Tchsp 18.3 reduced the oviposition amount by 94.7%. These results suggest that Tchsp 18.3 is not only essential for the resistance to heat and starvation stress, but also is critical for normal development and reproduction in T. castaneum.     

Crinkled employs wingless pathway for wing development in Tribolium castaneum

Crinkled is associated with embryonic denticle formation and auditory organ development in Drosophila melanogaster. However, the functions of Crinkled have not been fully investigated. Additionally, the genes that participate in the Crinkled pathway are unknown. Phylogenetic analysis indicates that crinkled exhibits a one‐to‐one orthologous relationship in insects. In Tribolium castaneum, the crinkled gene is 6,498 bp in length and consists of six exons. Crinkled expression peaked during two phases in Tribolium: late embryonic and pupal stages. High levels of crinkled mRNA were detected in the fat body, head, epidermis, ovary, and accessory gland of late adults. Knockdown of crinkled using RNA interference (RNAi) severely affected wing morphogenesis in T. castaneum. We further showed that crinkled silencing reduced forked expression through wingless and shaven‐baby, and RNAi of forked phenocopied the effects of crinkled knockdown in T. castaneum. This study investigated the development role of crinkled in postembryonic stages and indicated that forked mediates the functions of crinkled during wing morphogenesis in T. castaneum.     

First evidence of DNA methylation in insect Tribolium castaneum

DNA methylation has been studied in many eukaryotic organisms, in particular vertebrates, and was implicated in developmental and phenotypic variations. Little is known about the role of DNA methylation in invertebrates, although insects are considered as excellent models for studying the evolution of DNA methylation. In the red flour beetle, Tribolium castaneum (Tenebrionidae, Coleoptera), no evidence of DNA methylation has been found till now. In this paper, a cytosine methylation in Tribolium castaneum embryos was detected by methylation sensitive restriction endonucleases and immuno-dot blot assay. DNA methylation in embryos is followed by a global demethylation in larvae, pupae and adults. DNA demethylation seems to proceed actively through 5-hydroxymethylcytosine, most probably by the action of TET enzyme. Bisulfite sequencing of a highly abundant satellite DNA located in pericentromeric heterochromatin revealed similar profile of cytosine methylation in adults and embryos. Cytosine methylation was not only restricted to CpG sites but was found at CpA, CpT and CpC sites. In addition, complete cytosine demethylation of heterochromatic satellite DNA was induced by heat stress. The results reveal existence of DNA methylation cycling in T. castaneum ranging from strong overall cytosine methylation in embryos to a weak DNA methylation in other developmental stages. Nevertheless, DNA methylation is preserved within heterochromatin during development, indicating its role in heterochromatin formation and maintenance. It is, however, strongly affected by heat stress, suggesting a role for DNA methylation in heterochromatin structure modulation during heat stress response.