Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa (2n =2X = 14, VV), a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetie Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge (a remarkable morphology) mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V(2D) and the hybrid F1 was then self-crossed. Chromosome C-banding, genomie in situ hybridization (GISH), and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids F2 and F3. To date, four translocations including one small segmental translocation T6BS.6BL-2VS, two whole arm translocations (preliminarily designed as T3DS·2VL and T2VS·7DL) and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one iso- chromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25-120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 (2BL) tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS·7DL, Mt2VS, 2VS·2VS as well as the parent DS2V(2D), whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C of Aegilops triuncialis could successfully induce chromosome 2V structural changes.     

利用离果山羊草3C染色体诱导簇毛麦2V染色体结构变异

【目的】簇毛麦是普通小麦的一个近缘物种,它具有许多抗病基因,在小麦育种中起重要作用。抗白粉病基因Pm21已被南京农业大学细胞遗传所成功地转移到小麦背景中,并被广泛地用于小麦育种实践。为了进一步转移和利用定位于簇毛麦2V染色体上的有用基因,如抗眼斑病基因、抗条锈基因和护颖颖脊刚毛基因,为小麦育种创造新种质。【方法】通过普通小麦农林26-离果山羊草3C二体异附加系与小麦-簇毛麦2V（2D）二体代换系杂交,综合运用染色体C-分带、基因组原位杂交、染色体构型分析和分子标记分析。【结果】从杂种F２和F３中鉴定出涉及簇毛麦2V结构变异的异染色体系7份,包括纯合缺失系1份（Del 2VS•2VL-）,易位系4份,其中纯合易位2份（初步推断为T3DS•2VL,T2VS•7DL）、小片段易位1份（T6BS•6BL-2VS）和中间插入易位1份（T2VS•2VL-W-2VL）,等臂染色体1份（2VS•2VS）和单端体1份（Mt2VS）。利用可分别追踪2VS 和2VL的分子标记Xwmc25-120和NAU/STSBCD135-1进行PCR分析,进一步证明这7份异染色体系中涉及簇毛麦2V染色体片段。【结论】涉及2V短臂的单端体Mt2VS,等臂染色体2VS•2VS和易位系T2VS•7DL在护颖颖脊上有簇状分布的刚毛,而涉及2V长臂的易位系T3DS•2VL无刚毛,进一步证实簇毛麦护颖颖脊刚毛基因位于2VS。离果山羊草3C染色体可有效诱发簇毛麦2V染色体结构变异。     

Development of EST-PCR Markers for the Chromosome 4V of Haynaldia villosa and Their Application in Identification of 4V Chromosome Structural Aberrants

EST-PCR based molecular markers specific for alien chromosomes are not only useful for the detection of the introgressed alien chromatin in the wheat background, but also provide evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers on chromosome 4V of Haynaldia villosa, a total of 607 primer pairs were designed according to the EST sequences, which were previously located in 23 different bins of wheat chromosomes 4A, 4B and 4D. By using the Triticum durum-H, villosa amphiploid and T. aestivum-H, villosa alien chromosome lines involving chromosome 4V, it was found that 9.23% of the tested primers could amplify specific bands for chromosome 4V. Thirty and twenty-six specific markers could be assigned to chromosome arms 4VS and 4VL, respectively. These 4V specific markers provided efficient tools for the characterization of structural variation involving the chromosome 4V as well as for the selection of useful genes located on chromosome 4V in breeding programs.     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育(英文)

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育(英文)

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育

[目的]为进一步利用簇毛麦2V染色体上的有益基因,为小麦育种提供新种质。[方法]通过普通小麦-簇毛麦2V(2D)二体代换系(DS2V)与普通小麦农林26-离果山羊草3C染色体二体异附加系(DA3C)杂交,综合运用染色体C-分带、基因组原位杂交和分子标记分析,并结合性状调查。[结果]从杂种后代中选育出小麦-簇毛麦纯合易位系T6BS.6BL-2VS,性状调查发现该易位系植株护颖颖脊上有刚毛。[结论]该易位系为杀配子染色体诱发的小片段易位;簇毛麦护颖颖脊刚毛基因定位于2VS的中部至端部。     

雌雄克氏原螯虾染色体核型比较分析

克氏原螯虾（Procambarus clarkii）染色体数目众多、形态短小的特征,增大了染色体研究的难度。为了更深入地了解克氏原螯虾的细胞遗传学特征,采用一种改进的染色体制备方法,从克氏原螯虾的触角腺中获得了高质量的染色体中期相,并比较了雌雄克氏原螯虾染色体核型之间的差异。结果表明,雌雄克氏原螯虾在染色体数目、染色体配对及分组上是一致的,即雌雄克氏原螯虾的染色体数目均为2n=188,核型公式为n=94=61m+19sm+7st+7T。雌雄克氏原螯虾在染色体核型数据上有所差异,雌性克氏原螯虾的染色体相对长度范围为0.72%～1.61%,雄性克氏原螯虾的染色体相对长度范围为0.58%～1.89%;雌雄克氏原螯虾最大的染色体均是sm型,而雌雄最小的染色体不同型,雄性最小的染色体为T型,雌性最小的染色体为st型;雌雄克氏原螯虾的st型染色体的形态差别最大,但未发现异型性染色体。     

六倍体小黑麦染色体的荧光原位杂交分析

采用基因组原位杂交(GISH)和荧光原位杂交(FISH)技术对六倍体小黑麦进行了研究,以鉴定其含有的黑麦染色体的数目和结构特点,了解小黑麦基因组组成,为其在小麦遗传育种中的应用提供参考.分别以黑麦基因组DNA及pSc200和pSc250探针进行顺序GISH-FISH分析,结果显示该小黑麦含有21对染色体,来源于黑麦的2对染色体的长臂端部出现pSc200和pSc250信号,而另外5对染色体的两端部都有该信号.以Oligo-pSc119.2-1,pSc200和pSc250探针对小黑麦进行了双色FISH分析,发现小黑麦的Oligo-pSc119.2-1信号与黑麦基本相似,而pSc200和pSc250信号与黑麦的差异较大.以黑麦基因组DNA及Oligo-pAs1-1和Oligo-pSc119.2-1为探针对小黑麦进行顺序GISH-FISH分析,鉴定其基因组组成为AABBRR;同时发现小黑麦染色体的一些FISH信号发生了变化,其原因可能是小黑麦形成过程中染色体结构发生了改变,从而导致小黑麦的DNA重复序列呈现多态性.     

普通小麦-长穗偃麦草异附加系的分子标记鉴定

利用分布于小麦7个部分同源群的1 246对引物对15个可能的小麦-长穗偃麦草二体异附加系的4个亲本的基因组DNA进行扩增。结果表明,186对SSR、209对EST-SSR和22对STS引物能够在长穗偃麦草中扩增出不同于其他3个小麦亲本的特异条带,其中18对引物可以在14个附加系中的1~7个附加系扩增出长穗偃麦草的特异带,说明它们可能添加长穗偃麦草的遗传物质。5个附加系(1-3、1-8、1-27、2-6和2-22)可能含有长穗偃麦草第7同源群染色体,4个附加系(5-10、5-20、6-23和6-18)可能含有长穗偃麦草第6同源群染色体。附加系1-13可能附加2条不同同源群的长穗偃麦草染色体。同时发现,小麦与长穗偃麦草杂交及其后代衍生过程中长穗偃麦草染色体间可能发生遗传重组。     

两种弧菌感染大黄鱼免疫相关基因的SNP位点分析

为了探究大黄鱼(Larimichthys crocea)免疫相关基因的SNP与弧菌抗性关系,分别利用鳗弧菌(Vibrio anguillarum)和副溶血弧菌(Vibrio parahaemolyticus)人工感染大黄鱼。对感染前后抗感群体的转录组进行高通量测序、筛选并分析其抗病差异:(1)筛选氨基酸的非同义突变SNP位点在抗鳗弧菌组有17个,而抗副溶血弧菌组的有28个;(2)一代测序验证结果发现,染色体NW_011323507.1上白细胞介素6受体基因(IL-6R)第91 196位碱基G突变为C,导致缬氨酸突变为亮氨酸,该位点G/C在抗鳗弧菌组、对照组样本之间突变基因型CC频率分别为12.5%和0,呈显著性差异(P0.05);(3)补体C1q/肿瘤坏死因子相关蛋白9(CTRP9)基因在染色体NW_011323975.1上的35 665位碱基突变(A-G),在副溶血弧菌抗感易感群体中突变位点基因型GG频率分别为37.5%和0,呈极显著性差异(P0.01)。结果表明,IL-6R-91196-G/C位点突变与大黄鱼抗鳗弧菌有关联,CTRP9-35665-A/G位点突变与大黄鱼抗副溶血弧菌有关联,这为大黄鱼抗弧菌群体的选育提供了理论依据。     

普通小麦-簇毛麦2V染色体端体异附加系的选育与鉴定

簇毛麦(Haynaldia villosa) 具有抗多种病害、耐旱、耐寒等优良性状,是可供小麦改良利用的优良遗传资源.本研究综合利用根尖细胞有丝分裂中期染色体Giemsa C-分带、花粉母细胞减数分裂中期Ⅰ染色体构型分析、荧光原位杂交(genomic in situ hybridization,GISH) 及分子标记等技术,从普通小麦-簇毛麦2V(2D) 异代换系与含有Ph抑制基因的中国春高配对材料(pairing homoeologous inhibitor,Ph1) CO4-13的杂交后代中选了出分别含有簇毛麦2V长臂和短臂的普通小麦-簇毛麦2V端体异附加系.含有簇毛麦2V短臂的附加系在护颖颖脊上有刚毛,而在含2V长臂的附加系的护颖颖脊上没有刚毛,可将控制护颖颖脊刚毛性状的基因进一步定位在簇毛麦2V染色体的短臂上.筛选出町以追踪2V染色体短臂的SSR标记wmc25.该分子标记和护颖颖脊刚毛形态标记可用来追踪导入小麦遗传背景中的簇毛麦2V染色体短臂.     

Grain proteomic analysis reveals central stress responsive proteins caused by wheat-Haynaldia villosa 6VS/6AL translocation

Haynaldia villosa (2n=14, VV), a wild grass of the subtribe Triticeae, serves as potential gene resources for wheat genetic improvement. In this study, the proteome characterization during grain development of Yangmai 5 and Yangmai 5-H. villosa 6VS/6AL translocation line was investigated by a comparative proteomic approach. Two-dimensional electrophoresis identified 81 differentially accumulated proteins (DAPs) during five grain developmental stages in wheat-H. villosa translocation line. These proteins were mainly involved in stress defense, storage protein, energy metabolism, protein metabolism and folding, carbon metabolism, nitrogen metabolism, and starch metabolism. In particular, 6VS/6AL translocation led to significant upregulation of 36 DAPs and specific expression of 11 DAPs such as chitinase, thaumatin-like proteins, glutathione transferase, α-amylase inhibitor, heat shock proteins, and betaine aldehyde dehydrogenase. These proteins mainly involved in biotic and abiotic stress responses. Further analysis found that the upstream 1 500 promoter regions of these stress-responsive DAP genes contained multiple high-frequency cis-acting elements related to stress defense such as abscisic acid response element ABRE, methyl jasmonate (MeJA)-response element TGACG-motif and CGTCA-motif involved in oxidative stress and antioxidant response element (ARE). RNA-seq and RT-qPCR analyses revealed the high expression of these stress-defensive DAP genes in the developing grains, particularly at the early-middle grain filling stages. Our results demonstrated that 6VS chromosome of H. villosa contains abundant stress-defensive proteins that have potential values for wheat genetic improvement.     

Development of EST-PCR markers specific to the long arm of chromosome 6V of Dasypyrum villosum

Expressed sequence tags-derived polymerase chain reaction (EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6VL of Dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags (EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6AL, 6BL, and 6DL. By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6V, it was found that 32 (10.77%) primers could amplify specific bands for chromosome 6V, and 31 could be allocated to chromosome arm 6VL. These 6VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6VL in common wheat background as well as for the selection of useful genes located on 6VL in breeding programs.     

Molecular Mapping of Two Novel Stripe Rust Resistant Genes YrTp1 and YrTp2 in A-3 Derived from Triticum aestivum × Thinopyrum ponticum

Loss of variety resistance to stripe rust (Puccinia striiformis Westend f. sp.tritici) is an important factor causing massive periodical epidemic of rust in wheat production. Creation and development of new races of rust pathogen have led to serious crisis of resistance loss in widely planted varieties. This has quickened the search for new resistance resources. Molecular marker could facilitate the identification of the location of novel genes. A line A-3 with high resistance (immune) to currently epidemic yellow rust races (CY29, 31, 32) was screened out in offspring of Triticum aestivum × Thinopyrum ponticum. Segregation in F₂ and BC₁ populations indicated that the resistance was controlled by two independent genes: one dominant and one recessive. SSR markers were employed to map the two resistant genes in the F₂ and BC₁ populations. A marker WMC477-_167bp located on 2BS was linked to the dominant gene with genetic distance of 0.4 cM. Another marker WMC364-_{208 bp} located on 7BS was linked to the recessive-resistant gene with genetic distance of 5.8 cM. The two genes identified in this paper might be two novel stripe rust resistant genes, which were temporarily designated as YrTp1 and YrTp2, respectively. The tightly linking markers facilitate transfer of the two resistant genes into the new varieties to control epidemic of yellow rust.     

Inheritance and Molecular Mapping of Stripe Rust Resistance Gene Yr88375 in Chinese Wheat Line Zhongliang 88375

Stripe rust is one of the most important diseases of wheat worldwide. Inheritance of stripe rust resistance and mapping of resistance gene with simple sequence repeat （SSR） markers are studied to formulate efficient strategies for breeding cultivars resistant to stripe rust. Zhongliang 88375, a common wheat line, is highly resistant to all three rusts of wheat in China. The gene conferring rust disease was deduced originating from Elytrigia intermedium. Genetic analysis of Zhongliang 88375 indicated that the resistance to PST race CYR31 was controlled by a single dominant gene, temporarily designated as Yr88375. To molecular map Yr88375, a F2 segregating population consisting of 163 individuals was constructed on the basis of the hybridization between Zhongliang 88375 and a susceptible wheat line Mingxian 169; 320 SSR primer pairs were used for analyzing the genetic linkage relation. Six SSR markers, Xgwm335, Xwmc289, Xwmc810, Xgdmll6, Xbarc59, and Xwmc783, are linked to Yr88375 as they were all located on chromosome 5BL Yr88375 was also located on that chromosome arm, closely linked to Xgdmll6 and Xwmc810 with genetic distances of 3.1 and 3.9 cM, respectively. The furthest marker Xwmc783 was 13.5 cM to Yr88375. Hence, pedigree analysis of Zhongliang 88375 combined with SSR markers supports the conclusion that the highly resistance gene Yr88375 derived from Elytrigia intermedium is a novel gene for resistance to stripe rust in wheat. It could play an important role in wheat breeding programs for stripe rust resistance.     

小麦TaSIM基因结构及表达分析

根据小麦TaSIM基因的cDNA序列设计引物,采用PCR技术从小麦中克隆TaSIM基因的DNA序列,采用半定量RT-PCR方法研究TaSIM基因在不同组织中的表达。结果表明:TaSIM编码区DNA序列长度为2 355bp,包含2个外显子和1个内含子。分析发现内含子富含AT,在内含子中发现2个MYB转录因子结合位点。半定量RT-PCR检测表明,TaSIM基因在不同组织中均有表达,在雄蕊中的表达量最高。TaSIM表达量依次为雄蕊>雌蕊>根>茎>叶。     

山葡萄C4H基因的克隆表达及遗传转化分析

通过生物学技术来研究C4H基因在山葡萄着色过程中的作用,进而揭示山葡萄果皮着色的分子机理;利用RT-PCR技术克隆了山葡萄C4H基因的全长cDNA序列,并对该蛋白进行生物信息学分析,预测其功能;利用实时荧光定量PCR检测C4H基因在山葡萄8个不同转色时期的表达量,将克隆获得的山葡萄C4H基因完整的ORF连接到原核表达载体pET28a上,转化到大肠杆菌E.coli BL21(DE3),并通过不同浓度的IPTG诱导表达, SDS-PAGE检测表达产物.为了验证山葡萄C4H基因的功能,构建了表达载体pC C4H并转化农杆菌GV3101.用菌液浸泡花序法对拟南芥进行遗传转化,在含50 mg/L Kan的培养基上对T_0代种子进行筛选.克隆获得的山葡萄C4H cDNA全长1 735 bp,开放阅读框1 518 bp,编码505个氨基酸,该基因表达产物分子质量为57.70 KDa,等电点值9.06.C4H基因在山葡萄果皮转色各个时期均存在表达;该基因原核表达产物与预期大小一致,表明原核表达成功,拟南芥遗传转化先后得到3个阳性幼苗.对移栽成活的2株抗性植株进行PCR检测为阳性, 2株叶片颜色均变成紫红色;经花色素苷质量浓度的测定表明其质量浓度比对照组植株高出3倍.在拟南芥中花色素苷质量浓度虽然较低,但还是能少量合成,说明其花色素苷生物合成途径是开通的,只是积累的量较少.     

pH及盐分对大丽轮枝菌微菌核形成的影响

微菌核是棉花黄萎病原大丽轮枝菌(Verticillium dahliae Kleb.)在土壤中的主要存活形式及该病的侵染源,研究土壤pH及盐分对大丽轮枝菌微菌核形成的影响对明确黄萎病发生具有重要意义。采用菌丝生长速率法研究培养基pH及盐分质量浓度与种类对病原菌生长及微菌核形成的影响。供试大丽轮枝菌菌丝生长最适pH为7.0,偏酸或偏碱会抑制菌丝生长,但培养基偏碱可显著促进微菌核形成。当pH为8.0时,大丽轮枝菌菌丝生长受抑制较小,同时微菌核区面积较pH为7.0时增加22.6%。盐分质量浓度影响大丽轮枝菌菌丝生长及微菌核形成。随培养基NaCl质量浓度增加,供试大丽轮枝菌生长受到抑制,菌落面积和菌丝面积均逐渐减小,但微菌核形成量却显著增加;当NaCl质量浓度为10g·L~(-1)时,微菌核区面积较无NaCl时增加40.7%。盐分种类影响供试大丽轮枝菌生长。随盐分质量浓度增加,氯化物(NaCl和KCl)和硫酸盐(Na_2SO_4和MgSO_4)均可促进大丽轮枝菌微菌核形成,而CaCl_2则显著促进菌丝生长,并在质量浓度大于7g·L~(-1)时抑制微菌核形成。在培养环境偏碱性或氯化物和硫酸盐含盐量较高时,均可促进棉花黄萎病原大丽轮枝菌微菌核形成量增加。     

施加硒肥条件下的印度梨形孢接种对香榧幼苗根系发育、 气体交换及其叶片硒累积的影响

近些年来香榧果实在干果市场非常受到人们的喜爱,如何提高其果实产量,促进其果实品质尤其微量元素的强化是当前香榧栽培上需要解决的问题。微量元素硒是硒蛋白必需的元素,它与多种人体疾病有关。采用盆栽的方式,施加矿物硒粉,并对香榧幼苗接种根内生真菌印度梨形孢。结果表明,印度梨形孢可以侵染香榧幼苗的根系(侵染率为22.34%),并使其总根长、总根表面积和总根体积以及二级根上一级根的数量,分别增加12.69%、16.26%、20.81%和10.93%,但对其特定根长和根组织密度没有显著影响。香榧幼苗接种印度梨形孢可显著提高净光合速率、气孔导度、胞间二氧化碳浓度以及蒸腾速率,分别增加92.92%、6.16倍、47.67%和2.36倍,同时显著提高光能利用效率（88.62%）而降低水分利用效率（84.28%）。接种显著地促进了香榧幼苗总生物量的累积（36.94%）以及叶片总硒含量（1.68倍）。 基于印度梨形孢可促进植物营养吸收的特性,该根内生真菌可以用香榧硒的生物强化,以提高香榧果实的品质。     

基于线粒体Cytb序列的3个宽体金线蛭群体遗传多样性分析

对江苏省扬州高邮、苏州张家港、泰州海陵3个地区宽体金线蛭自然群体的线粒体Cytb基因全序列进行了测定和分析。结果显示:宽体金线蛭Cytb基因全长1 146 bp,样本的A、G、T、C平均含量分别为27.57%、15.77%、43.49%、13.17%,A+T含量(71.06%)明显高于G+C含量(28.94%)。在77个宽体金线蛭样本中,共检测到96个多态位点,其中有58个简约信息位点,获得47个单倍型,共享单倍型数目1个。3个宽体金线蛭群体的平均单倍型多样性、核苷酸多样性、平均核苷酸差异数分别为0.9781、0.0133、15.20,其中泰州海陵群体的单倍型多样度(0.987 7)和核苷酸多样度(0.013 1)最高。3个群体间的遗传分化指数F_(st)为0.065 1～0.175 6,且差异显著(P0.05),不同群体间存在一定的遗传分化。分子方差分析(AMOVA)表明,11.13%的变异来自群体间,88.87%的变异来自群体内。用邻接法构建的单倍型系统发育进化树显示,不同地理群体的个体呈交错分布,没有形成明显的地理谱系。中性检验和核酸不配对分布结果表明,3个宽体金线蛭野生群体总体大小保持相对稳定,没有发生明显的种群扩张。