病原性鳗弧菌(Vibrio anguillarum)双重PCR与LAMP检测方法的建立

本研究检测了分离自发病大菱鲆、半滑舌鳎及鲤鱼的22株病原鳗弧菌(Vibrio anguillarum)毒力相关基因的携带情况，并建立了病原鳗弧菌的分子生物学检测方法。以PCR方法检测8个毒力相关基因的分布，结果显示，22株病原鳗弧菌均可扩增出6个基因(empA、vah1、vah4、flaA、rtxA和tonB)目的条带，未扩增出virA和angM基因；针对vah4和rtxA设计引物进行双重PCR扩增，同一PCR反应体系可扩增出两条目的条带，灵敏度为2.4×103 CFU/ml，对照菌无任何扩增条带；以vah4设计引物进行LAMP扩增，病原鳗弧菌可扩增出阶梯状条带，呈现阳性反应，6株对照菌无阶梯状扩增条带且呈现阴性反应，LAMP扩增灵敏度为2.4×101 CFU/ml。LAMP检测灵敏度是双重PCR的100倍，LAMP技术与PCR比较，操作简便、快速、灵敏度高且不需昂贵仪器，LAMP检测鳗弧菌的方法更适合于养殖生产实际应用。     

病原性鳗弧菌(Vibrio anguillarum)双重PCR与LAMP检测方法的建立

本研究检测了分离自发病大菱鲆、半滑舌鳎及鲤鱼的22株病原鳗弧菌(Vibrio anguillarum)毒力相关基因的携带情况,并建立了病原鳗弧菌的分子生物学检测方法。以PCR方法检测8个毒力相关基因的分布,结果显示,22株病原鳗弧菌均可扩增出6个基因(empA、vah1、vah4、flaA、rtxA和tonB)目的条带,未扩增出virA和angM基因;针对vah4和rtxA设计引物进行双重PCR扩增,同一PCR反应体系可扩增出两条目的条带,灵敏度为2.4×103 CFU/ml,对照菌无任何扩增条带;以vah4设计引物进行LAMP扩增,病原鳗弧菌可扩增出阶梯状条带,呈现阳性反应,6株对照菌无阶梯状扩增条带且呈现阴性反应,LAMP扩增灵敏度为2.4×10~1 CFU/ml。LAMP检测灵敏度是双重PCR的100倍,LAMP技术与PCR比较,操作简便、快速、灵敏度高且不需昂贵仪器,LAMP检测鳗弧菌的方法更适合于养殖生产实际应用。     

迟钝爱德华菌环介导等温扩增检测方法研究

为了建立一套用于实验室及室外现场检测迟钝爱德华菌(Edwardsiella tarda, Et)的环介导等温扩增方法(loop-mediated isothermal amplification, LAMP)，以迟钝爱德华菌的毒力基因fimA为靶基因设计特异性引物，以基因组DNA为模板，进行环介导恒温扩增，并对其特异性、灵敏性和临床检测进行了试验。结果显示，迟钝爱德华菌阳性样本反应呈现为荧光绿色，阴性样本不变色。该LAMP方法的最适反应温度为63℃；特异性试验表明仅迟钝爱德华菌样本发生反应，而杀鲑气单胞菌、嗜水气单胞菌、温气单胞菌和豚鼠气单胞菌均不发生反应；敏感性试验表明，该LAMP方法可检出浓度为2.16×10^-5mg/L的迟钝爱德华菌的核酸。     

二温式聚合酶链反应鉴别诊断迟缓爱德华氏菌病

根据迟缓爱德华氏菌株23S rDNA基因,设计合成一对引物XZE15、XZE16,建立了二温式聚合酶链反应(PER)鉴别诊断迟缓爱德华氏菌株的技术.特异性试验表明,对3株迟钝爱德华氏菌株进行PCR扩增出与预期大小相一致的284 bp的特异性片段,但对3株钻鱼爱德华氏菌及其他对照鱼病病原体核酸模板的PCR扩增不出现任何条带;敏感性试验结果显示,该二温式PCR可以检测到10 Pg的迟钝爱德华氏菌DNA.     

黑棘鲷内脏结节病病原的分离、鉴定及致病性研究

2017年4月,浙江台州某海水养殖公司跑道式养殖池黑棘鲷大量发病,病鱼活力下降、食欲减退、体表溃疡,剖检可见肝脏、脾脏、肾脏肿大且有不同程度的白色结节,同时伴随大量腹水。用胰蛋白胨大豆琼脂(TSA)从典型结节病濒死黑棘鲷器官分离到革兰阴性短杆菌。分离病原菌AS15对健康黑棘鲷的致病力结果显示,AS15腹腔注射可使健康黑棘鲷发病死亡,死亡黑棘鲷可出现自然发病症状,在(24±1)°C的条件下,(25±2) g黑棘鲷的半数致死浓度(LD_(50))为6.5×10~4 CFU/尾。经API 20E鉴定,分离菌株AS15生理生化特性与类杀鱼爱德华氏菌LADL05-105和迟缓爱德华氏菌典型菌ATCC15947相似度为86.2%,与杀鱼爱德华氏菌ET~T883的相似度为82.8%;AS15的16S rDNA与杀鱼爱德华氏菌ET~T883同源性达99%,gyrB与类杀鱼爱德华氏菌LADL05-105和杀鱼爱德华氏菌ETT883同源性分别为100%和98%;16S rDNA进化树显示,AS15与ETT883、LADL05-105聚为一簇,gyrB进化树与LADL05-105、NCIM2056聚为一簇;采用4种爱德华氏菌属种特异性引物对AS15进行PCR分析,结果显示,AS15可扩增出类杀鱼爱德华氏菌的种特异性片段,不能扩增出迟缓爱德华氏菌、鲇鱼爱德华氏菌、杀鱼爱德华氏菌的种特异性片段,表明AS15属类杀鱼爱德华氏菌成员。分析了AS15的菌毛基因、sodB等毒力基因,发现AS15具有fimA、fimB、fimC、fimD等4种菌毛基因和sodB、citC、esrB、mukF、katB等毒力基因。本实验首次从黑棘鲷上检出致病性类杀鱼爱德华氏菌,该菌对黑棘鲷的发病机制和毒力机理还需进一步研究。     

5种淡水细菌的多重PCR检测方法的建立及其应用

细菌对水产养殖品种危害大,造成的经济损失高。传统的显微镜镜检方法检测细菌误差大,而利用PCR方法鉴定细菌种类特异性强,准确性高。利用多重PCR探索出一种能准确鉴定水产常见细菌。研究表明,利用PCR分别扩增了嗜水气单胞菌Hly A基因,肺炎克雷伯菌Khe基因,无乳链球菌Sip基因,爱德华氏菌Ser C基因和Eip基因以及迟钝爱德华氏菌Muk F基因和Gad B基因,扩增特异性强。并成功地建立了一种同时用于无乳链球菌和爱德华氏菌等两种细菌以及爱德华氏菌、肺炎克雷伯菌、迟钝爱德华氏菌和嗜水气单胞菌等4种细菌的多重PCR检测方法。为了验证多重PCR方法的准确性,采集患病黄鳝病灶部位,分离培养细菌并提取该细菌基因组DNA,用上述两种多重PCR方法检测出患病黄鳝体内感染的细菌主要是肺炎克雷伯菌和嗜水气单胞菌。建立的多重PCR检测方法对水产常见致病菌的快速诊断和分子流行病学的调查具有重要意义,为鱼类的疾病诊断提供了依据。     

一株黄颡鱼源迟缓爱德华氏菌弱毒株的分离鉴定及特性分析

为探明湖北荆州市某养殖基地患病黄颡鱼(Pelteobagrus fulvidraco)以烂身、腹水为主要症状且持续性死亡的确切病因，本研究对患病黄颡鱼进行了病原筛查，从患病黄颡鱼组织中分离到一株优势菌，经过形态特征、生理生化鉴定、16S rRNA测序和系统进化分析确定分离菌株为迟缓爱德华氏菌，命名为Et-4。结果显示：分离菌株Et-4具有较强的生物被膜形成能力，不携带esaV、fimA、gadB和katB四种主要的毒力基因。分离菌株Et-4人工感染健康黄颡鱼出现与自然发病类似的症状，并能从病鱼中再次分离出该菌，证实迟缓爱德华氏菌是引起此次黄颡鱼持续性死亡的致病原；分离菌株Et-4对黄颡鱼的LD₅₀为3.9×10⁶ CFU/g,结合毒力基因谱判定其为弱毒株。分离株Et-4携带耐药基因tet A、sulⅠ和add A1,对头孢类药物、庆大霉素、新霉素等敏感；对四环素类、青霉素类、万古菌素等耐药；中药抑菌实验表明乌梅和丁香对迟缓爱德华氏菌Et-4有明显的抑制效果。     

龟鲍曼不动杆菌和摩氏摩根菌双重PCR诊断方法建立

为建立龟源致病性鲍曼不动杆菌和摩氏摩根菌双重PCR诊断方法,应用鲍曼不动杆菌rpoB基因和摩氏摩根菌gyrB基因的特异性引物,对单重和双重PCR反应的退火温度、引物浓度、MgCl_2浓度、dNTP浓度等进行优化。试验结果显示,鲍曼不动杆菌rpoB基因和摩氏摩根菌gyrB基因引物分别进行PCR扩增后,均获得特异性条带,与预期结果一致;rpoB和gyrB基因的上、下游引物各为0.3μmol/L,退火温度为59.3℃,dNTP浓度为0.5 mmol/L,MgCl_2浓度为5 mmol/L时,PCR扩增效果最佳;特异性试验结果显示,本方法仅对摩氏摩根菌和鲍曼不动杆菌有扩增,对其他病原菌如肺炎克雷伯氏菌、维氏气单胞菌及恶臭假单胞菌等无扩增;灵敏性结果表明,本方法对鲍曼不动杆菌和摩氏摩根菌最低检测质量浓度分别为4.12×10~(-3) ng/μL和1.257×10~(-3) ng/μL。通过对临床病料样本的检测分析证明,本方法与单重PCR检测结果一致。本试验所建立的针对两种病原菌检测的双重PCR方法特异性强、灵敏度高,可为龟鳖类鲍曼不动杆菌和摩氏摩根菌的鉴别诊断和临床调查提供帮助。     

基于toxR基因病原哈氏弧菌PCR检测方法的建立

基于toxR基因序列设计检测哈氏弧菌的1对特异性引物,通过PCR反应条件优化、PCR反应特异性及敏感性检测,建立一种哈氏弧菌的特异性分子检测方法.试验结果表明,该引物可使哈氏弧菌扩增出大小为390 bp的toxR基因片段,3种水产动物病原弧菌未扩增出任何条带,敏感性检测结果为该PCR反应最低能检测出0.375 ng/μ...     

牙鲆迟钝爱德华氏菌血清型及荧光抗体检验

以迟钝爱德华氏菌的代表菌株(HC010907-1)为免疫原,制备免疫血清,对分离于牙鲆鱼的130株迟钝爱德华氏菌进行了血清型检定,结果表明:供试的130株迟钝爱德华氏菌均为同种血清型;同时以此免疫血清为第一抗体,以标准羊抗兔IgG荧光抗体为第二抗体,进行了荧光抗体技术检验迟钝爱德华氏菌的可行性试验,表明亦具有较强的特异性。     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Characteristics of the marine environment and algal blooms in Gamak Bay

This study was conducted to gain insight into the characteristics of algal blooms in relation to the marine environment of Gamak Bay. From the first known occurrence of algal blooms in 1984 until 2006, 23 causal species have been identified, the most common ones being Prorocentrum sp., Cochlodinium polykrikoides, Chaetoceros sp., Skeletonema costatum, and Heterosigma akashiwo. A principal component analysis indicated that blooms of these species develop under different conditions in terms of water temperature, salinity, precipitation, and insolation. Field data showed Cochlodinium polykrikoides to be favored by high temperatures and to be euryhaline, whereas Skeletonema costatum appeared to be eurythermal and euryhaline. Prorocentrum sp. and Chaetoceros sp. appeared to be stenothermal and stenohaline. Finally, Heterosigma akashiwo appeared at the lowest temperature and highest salinity of the five species and was classified here as stenothermal and stenohaline.     

Bioaugmentation in the growth and water quality of livebearing ornamental fishes

Bacillus subtilis isolated from the intestine of Cirrhinus mrigala (Hamilton) was incorporated into the rearing water of Poecilia reticulata (Peters), Poecilia sphenops (Valenciennes), Xiphophorus helleri (Heckel) and Xiphophorus maculatus (Gunther) at four different concentrations (5 × 10⁸ cells ml⁻¹, 5 × 10⁷ cells ml⁻¹, 5 × 10⁶ cells ml⁻¹ and 5 × 10⁵ cells ml⁻¹) and its effect on fish growth performance and survival, water quality parameters and bacterial population of water were assessed. The results showed that the addition of bacterial cells in the rearing water resulted in greater survival and a faster growth rate and, hence, greater length and weight increments of the livebearers. The use of a bioaugmentor in the rearing water of the livebearing fishes resulted in significantly lower (P < 0.05) concentrations of dissolved organic matter and total ammonium nitrogen. The counts of motile aeromonads and total coliforms recorded in the water of bioaugmented tanks were also lower than that in the control tank. Bioaugmentation between 10⁶ and 10⁸ cells ml⁻¹ in the rearing water is sufficient in establishing a bioaugmentor and the use of a higher concentration of bacterial cells did not always lead to significantly better results.     

Use of Microbacterium sp. and Exiguobacterium mexicanum to improve the survival and development of Artemia under xenic conditions

The effect of Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N was evaluated in the diet of Artemia under xenic conditions. Viable cultures of bacteria were provided to xenic cultures of Artemia in combination with Sacharomyces cerevisiae, cornflour or Spirulina, and the effect on the survival and growth was recorded. The use of these bacterial strains improves significantly the survival of Artemia independently of the used food (P < 0.05), and variable results were observed in the growth.     

养殖大菱鲆中牙鲆肠弧菌的分离鉴定及组织病理学

2007年1月，山东省胶南某养殖场人工养殖的大菱鲆（Scophthalmus maximus）发生严重病害并大批死亡。病鱼的主要症状是体表溃疡，腹腔积液，肠道肿胀，肝脏萎缩，胆囊暗绿色等。从病鱼胆囊中分离纯化得到优势菌株，命名为da3。人工感染试验证实，该菌株对大菱鲆有较强的致病性。对体重为25 g的大菱鲆的半数致死量为每尾鱼2×106 cfu。通过细菌16S rDNA序列测定及形态学和生理生化特征研究确定，该病原菌为牙鲆肠弧菌（Vibrio ichthyoenteri）。组织病理学观察表明，病鱼的肝脏、肾脏、脾脏、肠道和脑的微观结构发生了明显的病理变化，由此引起的器官功能衰竭可能是病鱼死亡的主要原因。本文结果对大菱鲆细菌性病害的控制具有参考价值。     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.     

南海北部陆架水域多齿蛇鲻与花斑蛇鲻的年龄与生长

2014年7月至2015年5月采集南海北部陆架海域的多齿蛇鲻(Sauridatumbil)和花斑蛇鲻(Sauridaundosquamis)样本,测量其叉长、体重、性腺成熟度,并读取其耳石和日龄数据,对这2个种的生物学特征进行比较,发现多齿蛇鲻和花斑蛇鲻叉长、体重雌雄间差异显著(P0.05)。多齿蛇鲻雌性与雄性样本,叉长范围分别为109～310 mm、117～283 mm,体重范围分别为13～358 g、20～297 g,日龄范围分别为76～558 d、73～526 d, von Bertalanffy生长方程的相关参数分别为:L_(∞♀)=278 mm, L_(∞♂)=276 mm; K_♀=1.30, K_♂=1.30; t_(0♀)=-0.07 a, t_(0♂)=-0.08 a。雌性性腺成熟度Ⅲ期样本从350～400 d日龄组开始出现,雌性与雄性样本孵化高峰期在3月、7月、9—10月。花斑蛇鲻雌性与雄性样本,叉长范围分别为105～269 mm、95～264 mm,体重范围分别为11～220 g、8～228 g,日龄范围分别为79～486 d、84～488 d, von Bertalanffy生长方程的相关参数分别为:L_(∞♀)=269 mm, L_(∞♂)=265 mm, K_♀=1.29, K_♂=1.29, t_(0♀)=-0.06 a, t_(0♂)=-0.06 a,雌性性腺成熟度Ⅲ期样本从200～250 d日龄组开始出现,雌性与雄性样本孵化高峰期在6—11月。与历史资料进行对比,发现南海海域多齿蛇鲻与花斑蛇鲻出现个体大小逐渐减小、性成熟提前等适应性特征。结论认为,为了南海多齿蛇鲻与花斑蛇鲻渔业资源的可持续发展,需要降低南海近海的捕捞压力以促进南海渔业资源的发展。     

基于线粒体COⅠ和Cytb基因序列的北太平洋柔鱼种群遗传结构研究

为准确掌握柔鱼的种群遗传结构,拟通过线粒体DNA的COI和Cytb基因序列分析方法对柔鱼不同产卵季节群体的遗传多样性和遗传结构进行研究。经PCR扩增与测序分别获得600 bp COI与481 bp Cytb基因序列。基于COI基因序列分析得到的单倍型数、单倍型多样性指数、核苷酸多样性指数及平均核苷酸差异数分别为24、(0.729 ±0.033)、(0.005 70 ± 0.003 25)和3.421。基于Cytb基因序列分析得到的单倍型数、单倍型多样性指数、核苷酸多样性指数及平均核苷酸差异数分别为28、(0.852 ± 0.016)、(0.006 45 ± 0.003 73)和3.101。分析认为, 北太平洋柔鱼2个产卵季节群体均具有较高的单倍型多样性指数和较低的核苷酸多样性指数。单倍型邻接树、两两群体间的F_st值以及AMOVA分析结果均表明, 北太平洋柔鱼2个产卵季节群体间的遗传差异不显著, 不存在显著的群体遗传结构。初步认为, 该海域因缺乏地理上的障碍, 加之北太平洋海流的作用以及柔鱼个体较强的游泳能力, 使得群体之间具有较强的基因流。     

长江鲟类资源现状及保护

归纳了白鲟、长江鲟、中华鲟近40年自然资源量变化情况，统计了长江鲟和中华鲟人工增殖放流数据，评估了增殖放流成效。因物种特性和分布的差异，放流无效的原因迥异。过度捕捞使人工放流长江鲟在放流后6个月之内难逃被“误捕”，而中华鲟放流数量不足及长江和近海过度捕捞导致中华鲟人工增殖放流的贡献甚微，其结果是长江鲟和中华鲟增殖放流均无法达到自然繁殖群体的补充水平。本文探究了近40年来白鲟、中华鲟和长江鲟在保护、管理和决策上存在的误区和不足，提出在生态大保护的背景下，(1)应编制和实施《长江鲟拯救行动计划》优先项目；(2)应以《中华鲟拯救行动计划》为指引，在就地保护、迁地保护和遗传多样性保护方面提出更具有可操作性的优先计划；(3)应设立中华鲟和长江鲟物种拯救行动计划专项，对现有涉栖息地或保护区生态补偿项目进行优化整合。唯有以恢复长江鲟和中华鲟自然繁殖为核心，才有望延续和恢复其自然种群，实现人与自然的和谐共处，推动长江流域社会经济绿色可持续发展。     

三疣梭子蟹甲基转移酶2基因克隆及在胚胎、幼体和性腺发育过程中的表达分析

本研究采用SMART RACE方法克隆了三疣梭子蟹(Portunus trituberculatus)Dnmt2(Pt Dnmt2)基因。该基因c DNA全长为1291 bp,开放阅读框为1203 bp,编码400个氨基酸。结构域分析显示,PtDnmt2基因有典型的C5-DNA甲基化酶结构域。同源分析表明,PtDnmt2氨基酸序列与其他物种有较高的同源性。系统进化分析显示,三疣梭子蟹PtDnmt2氨基酸序列与罗氏沼虾(Macrobrachium rosenbergii)Dnmt2关系最近。qRT-PCR结果显示,PtDnmt2基因在三疣梭子蟹所有组织中均有表达,在卵巢中表达量显著高于其他组织(P0.05)。PtDnmt2基因在胚胎和幼体发育过程中的表达量随发育时期而变化,其在受精卵和多细胞时期无表达,从囊胚期开始出现,随着胚胎的发育表达量逐渐上升。在性腺发育不同时期PtDnmt2基因表达量存在显著差异,其在卵巢II期表达量最高,之后逐渐下降;在精巢中该基因的表达量随着精巢的发育逐渐上升,在IV期达到峰值。本研究结果表明,PtDnmt2基因参与了三疣梭子蟹胚胎、幼体和性腺发育调控。