Reproduction and breeding最新文献

Impact of photoperiod and temperature on melatonin, growth hormone, estradiol, and vitellogenin levels in female African catfish (Clarias gariepinus) reproduction cycle

Reproduction and breeding

Pub Date : 2025-01-24 DOI: 10.1016/j.repbre.2025.01.002

Epro Barades , Iskandar , Ibnu Dwi Buwono , Yuli Andriani

This study aimed to investigate the hormone levels of melatonin (Mel), estradiol (E2), vitellogenin (VTG), and growth hormone (GH) in inducing reproduction cycles in female African catfish, Clarias gariepinus, to develop actionable strategies that directly contribute to the efficiency and sustainability of African catfish farming. The treatments involved in this study are three photoperiod variations (L8:D16, L4:D20, and L0:D24) combined with two different temperatures (28 °C and 32 °C) during 90 days of culture (doc). Serum hormone levels were measured using ELISA, and egg diameter was measured using a microscope every 30 days. The results showed that the biological rhythm of the reproductive cycle of African catfish was accelerated by constant exposure to a photoperiod of L0:D24 and 28 °C for 30 days. In this condition, the levels of hormones involved in the reproductive such as Mel (89.82 ± 5.49 ng/mL), E2 (1.66 ± 0.02 ng/mL), VTG (100.96 ± 0.27 ng/mL) and GH (0.33 ± 0.02 ng/mL) with an average egg diameter of 1.15 ± 0.07 mm. These results highlight the complex interplay between photoperiod, temperature, and reproductive physiology in African catfish, suggesting that environmental manipulation could be a valuable tool for optimizing breeding conditions in aquaculture. The conclusion of this study is that manipulating the photoperiod and temperature is an effective and economical approach to stimulate fish spawning. These findings have important implications for African catfish breeding practices, as they provide a clear and actionable strategy for improving reproductive performance.

{"title":"Impact of photoperiod and temperature on melatonin, growth hormone, estradiol, and vitellogenin levels in female African catfish (Clarias gariepinus) reproduction cycle","authors":"Epro Barades , Iskandar , Ibnu Dwi Buwono , Yuli Andriani","doi":"10.1016/j.repbre.2025.01.002","DOIUrl":"10.1016/j.repbre.2025.01.002","url":null,"abstract":"<div><div>This study aimed to investigate the hormone levels of melatonin (Mel), estradiol (E2), vitellogenin (VTG), and growth hormone (GH) in inducing reproduction cycles in female African catfish, <em>Clarias gariepinus</em>, to develop actionable strategies that directly contribute to the efficiency and sustainability of African catfish farming. The treatments involved in this study are three photoperiod variations (L8:D16, L4:D20, and L0:D24) combined with two different temperatures (28 °C and 32 °C) during 90 days of culture (doc). Serum hormone levels were measured using ELISA, and egg diameter was measured using a microscope every 30 days. The results showed that the biological rhythm of the reproductive cycle of African catfish was accelerated by constant exposure to a photoperiod of L0:D24 and 28 °C for 30 days. In this condition, the levels of hormones involved in the reproductive such as Mel (89.82 ± 5.49 ng/mL), E2 (1.66 ± 0.02 ng/mL), VTG (100.96 ± 0.27 ng/mL) and GH (0.33 ± 0.02 ng/mL) with an average egg diameter of 1.15 ± 0.07 mm. These results highlight the complex interplay between photoperiod, temperature, and reproductive physiology in African catfish, suggesting that environmental manipulation could be a valuable tool for optimizing breeding conditions in aquaculture. The conclusion of this study is that manipulating the photoperiod and temperature is an effective and economical approach to stimulate fish spawning. These findings have important implications for African catfish breeding practices, as they provide a clear and actionable strategy for improving reproductive performance.</div></div>","PeriodicalId":74667,"journal":{"name":"Reproduction and breeding","volume":"5 1","pages":"Pages 54-66"},"PeriodicalIF":0.0,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143175461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Transcriptomic profiling reveals the mechanism of fast growth of an allodiploid hybrid fish

Reproduction and breeding

Pub Date : 2025-01-24 DOI: 10.1016/j.repbre.2025.01.001

Ziyi Huang , Siyang Huang , Hongwen Liu , Bei Li , Jianming Yu , Xuanyi Zhang , Fanglei Liu , Lujiao Duan , Qingfeng Liu

Distant hybridization is an important technique and widely used in fish genetic breeding. In previous research, we obtained the allodiploid hybrid (BR, 2n = 49) derived from the inter-subfamily hybridization of female blunt snout bream (Megalobrama amblycephala, BSB, 2n = 48) and male rare gudgeon (Gobiocypris rarus, RG, 2n = 50), which had obvious growth advantages over the male parent RG. However, less is known about the potential mechanisms of fast growth in BR. In this study, the liver transcriptomes of BR and parents were analyzed by RNA-seq technology. A total of 4.73 × 10⁸ clean reads and 11,040 co-expressed unigenes were obtained. The results showed that the differentially expressed genes in BR were asymmetric compared with those in both parents. Interestingly, the genome-wide expression level dominance (ELD) was biased toward the BSB genome in BR, even if BR inherited one more chromosome from RG than that from BSB. The KEGG analysis of ELD genes showed that most metabolism pathways were enriched in ELD-BSB, such as biosynthesis of unsaturated fatty acids, fatty acid elongation, pyruvate metabolism, and so on. Furthermore, the growth genes of BR were most enriched in the ELD-BSB (34.38 %) than in other categories. The expression levels of some key hormone genes (ghra, ghrb, igf1, igfals, igf2bp2, and grb2) were upregulated in BR compared with those in RG. This study provided an insight into the potential molecular mechanism of the fast growth in BR, which will be of great significance in fish genetic breeding.

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引用次数: 0

Cloning, expression, and localization of Tekt1 in sterile allotriploid crucian carp

Reproduction and breeding

Pub Date : 2025-01-08 DOI: 10.1016/j.repbre.2024.12.002

Shuxin Zhang , Liran Zhang , Faxian Yu, Xinge Ouyang, Haoxiang Yang, Qining Zuo, Yujie Huang, Xin Chen, Shengnan Li, Min Tao

Tektins (TEKTs) are constitutive proteins of microtubules associated with flagella, cilia, basal bodies, and centrioles. As one of the testis-specific candidate markers, Tekt1, the first identified member of the TEKT family in mammals, is intimately linked to the formation of sperm flagella and may play a pivotal role in flagellar stability and sperm motility. However, studies on Tekt1 in fish species are still relatively understudied. In this study, the full-length cDNAs of Tekt1 were respectively 1727 bp and 1696 bp in allotriploid crucian carp and diploid red crucian carp, which both comprised a 1209 bp open reading frame (ORF) encoding 402 amino acids. Conversely, the diploid common carp possessed a cDNA length of 1771 bp, characterized by a 1218 bp ORF encoding 405 amino acids. The Western blot analysis revealed that the expression level of Tekt1 protein in the testes of sterile allotriploid crucian carp was markedly decreased in comparison to that of fertile diploid red crucian carp during both pre-spermiation and spermiation periods. The immunohistochemistry analysis revealed abnormalities in the spermiogenesis of allotriploid crucian carp, showcasing a distinctive localization pattern of Tekt1 exclusively present in spermatids, in contrast to diploid red crucian carp, where Tekt1 was detected in both spermatids and spermatozoa. Taken together, these findings suggested differential expression of Tekt1 during spermiogenesis between allotriploid and diploid species, and indicated that the decreased expression of Tekt1 protein in allotriploid crucian carp might be associated with male sterility. Furthermore, these results pave the way for further exploration of reproductive characteristics in male allotriploid crucian carp and offer a theoretical foundation for future research on polyploid breeding.

{"title":"Cloning, expression, and localization of Tekt1 in sterile allotriploid crucian carp","authors":"Shuxin Zhang , Liran Zhang , Faxian Yu, Xinge Ouyang, Haoxiang Yang, Qining Zuo, Yujie Huang, Xin Chen, Shengnan Li, Min Tao","doi":"10.1016/j.repbre.2024.12.002","DOIUrl":"10.1016/j.repbre.2024.12.002","url":null,"abstract":"<div><div>Tektins (TEKTs) are constitutive proteins of microtubules associated with flagella, cilia, basal bodies, and centrioles. As one of the testis-specific candidate markers, <em>Tekt</em>1, the first identified member of the TEKT family in mammals, is intimately linked to the formation of sperm flagella and may play a pivotal role in flagellar stability and sperm motility. However, studies on <em>Tekt</em>1 in fish species are still relatively understudied. In this study, the full-length cDNAs of <em>Tekt</em>1 were respectively 1727 bp and 1696 bp in allotriploid crucian carp and diploid red crucian carp, which both comprised a 1209 bp open reading frame (ORF) encoding 402 amino acids. Conversely, the diploid common carp possessed a cDNA length of 1771 bp, characterized by a 1218 bp ORF encoding 405 amino acids. The Western blot analysis revealed that the expression level of Tekt1 protein in the testes of sterile allotriploid crucian carp was markedly decreased in comparison to that of fertile diploid red crucian carp during both pre-spermiation and spermiation periods. The immunohistochemistry analysis revealed abnormalities in the spermiogenesis of allotriploid crucian carp, showcasing a distinctive localization pattern of Tekt1 exclusively present in spermatids, in contrast to diploid red crucian carp, where Tekt1 was detected in both spermatids and spermatozoa. Taken together, these findings suggested differential expression of Tekt1 during spermiogenesis between allotriploid and diploid species, and indicated that the decreased expression of Tekt1 protein in allotriploid crucian carp might be associated with male sterility. Furthermore, these results pave the way for further exploration of reproductive characteristics in male allotriploid crucian carp and offer a theoretical foundation for future research on polyploid breeding.</div></div>","PeriodicalId":74667,"journal":{"name":"Reproduction and breeding","volume":"5 1","pages":"Pages 45-53"},"PeriodicalIF":0.0,"publicationDate":"2025-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143175463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Highly efficient disruption of tyrb gene using CRISPR/Cas9 in grass carp (Ctenopharyngodon idella)

Reproduction and breeding

Pub Date : 2025-01-04 DOI: 10.1016/j.repbre.2024.12.003

Pengfei Zhao , Jiaxiang Cheng , Liang Zhang , Wenbo Li, Shengfei Dai, Minghui Li, Deshou Wang, Xingyong Liu

Grass carp (Ctenopharyngodon idella) is the most important economic freshwater fish species in China. The stable production of high-quality grass carp depends significantly on excellent germplasm. In recent years, the generation of new germplasm based on genome editing has been applied to various cultured fish species. However, until now, there has been very few reports on the application of genome editing technology in grass carp. In this study, one-cell-stage embryos of grass carp were acquired through hormone-induced artificial spawning, thereby enabling the performance of genome editing in this species. The tyrb gene was isolated and chosen as the target of CRISPR/Cas9, because of its easily observable phenotype in F0 mutants. RT-PCR results indicated a high expression level of the tyrb gene in both skin and fin tissues. Subsequently, after the microinjection of the guide RNA (gRNA) and Cas9 protein mixture, targeted mutations were successfully identified through Sanger sequencing. Phenotypic analysis of the F0 mutants revealed that the disruption of tyrb led to a distinct golden phenotype, accompanied by a reduction or even absence of melanophores. Moreover, our data demonstrated that the combined utilization of two or three gRNAs caused large DNA fragment loss and a higher mutation rate in the F0 generation. Overall, this represents an application of CRISPR/Cas9 genome-editing technology in grass carp and may hold great significance for the future generation of new golden grass carp germplasm.

{"title":"Highly efficient disruption of tyrb gene using CRISPR/Cas9 in grass carp (Ctenopharyngodon idella)","authors":"Pengfei Zhao , Jiaxiang Cheng , Liang Zhang , Wenbo Li, Shengfei Dai, Minghui Li, Deshou Wang, Xingyong Liu","doi":"10.1016/j.repbre.2024.12.003","DOIUrl":"10.1016/j.repbre.2024.12.003","url":null,"abstract":"<div><div>Grass carp (<em>Ctenopharyngodon idella</em>) is the most important economic freshwater fish species in China. The stable production of high-quality grass carp depends significantly on excellent germplasm. In recent years, the generation of new germplasm based on genome editing has been applied to various cultured fish species. However, until now, there has been very few reports on the application of genome editing technology in grass carp. In this study, one-cell-stage embryos of grass carp were acquired through hormone-induced artificial spawning, thereby enabling the performance of genome editing in this species. The <em>tyrb</em> gene was isolated and chosen as the target of CRISPR/Cas9, because of its easily observable phenotype in F0 mutants. RT-PCR results indicated a high expression level of the <em>tyrb</em> gene in both skin and fin tissues. Subsequently, after the microinjection of the guide RNA (gRNA) and Cas9 protein mixture, targeted mutations were successfully identified through Sanger sequencing. Phenotypic analysis of the F0 mutants revealed that the disruption of <em>tyrb</em> led to a distinct golden phenotype, accompanied by a reduction or even absence of melanophores. Moreover, our data demonstrated that the combined utilization of two or three gRNAs caused large DNA fragment loss and a higher mutation rate in the F0 generation. Overall, this represents an application of CRISPR/Cas9 genome-editing technology in grass carp and may hold great significance for the future generation of new golden grass carp germplasm.</div></div>","PeriodicalId":74667,"journal":{"name":"Reproduction and breeding","volume":"5 1","pages":"Pages 38-44"},"PeriodicalIF":0.0,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143175460","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Fabrication and characterization of single-wall carbon nanotube and its biocompatibility to human hepatocytes

Reproduction and breeding

Pub Date : 2024-12-19 DOI: 10.1016/j.repbre.2024.12.001

Hatef Rahim Sabbaghizadeh , Arshin Oskoueian , Amir Hossein Ashtari

Single-wall carbon nanotubes (SWCNTs) have emerged as promising nanocarriers for targeted cancer drug delivery due to their unique structural properties. However, their cytotoxicity remains a significant challenge, as the biocompatibility of SWCNTs with human cells, particularly hepatocytes, is crucial for their clinical application. The toxicity of SWCNTs is influenced by factors such as nanoparticle size, morphology, surface chemistry, and the presence of impurities. In this study, we aimed to synthesize highly pure SWCNTs and assess their biocompatibility with human hepatocyte cells.

SWCNTs were fabricated using a modified chemical vapor deposition (CVD) method, followed by a two-step acid purification technique. Raman spectroscopy and electron microscopy confirmed a high purity level of 99.8 %. The biocompatibility of the purified SWCNTs was evaluated using an in vitro model with human hepatocytes. Results indicated that high concentrations of SWCNTs (>50 μg/ml) significantly reduced cell viability, increased lactate dehydrogenase leakage, and elevated lipid peroxidation, while simultaneously suppressing antioxidant enzyme activity.

Flow cytometry analysis further revealed that exposure to high concentrations of SWCNTs induced apoptosis in hepatocytes. Molecular analysis of key biomarkers demonstrated upregulation of TNF-α, IL1β, NF-kB, and iNOS, alongside downregulation of nrf2 gene and protein expression. These alterations contribute to the mechanisms underlying SWCNT-induced oxidative stress and apoptosis in human hepatocyte cells. Despite the high purity of SWCNTs, their cytotoxic effects may be attributed to their inherent physical properties, including rigidity, surface area, and fiber length.

In conclusion, while SWCNTs hold great potential for cancer drug delivery, managing their toxicity remains critical for their future therapeutic applications.

{"title":"Fabrication and characterization of single-wall carbon nanotube and its biocompatibility to human hepatocytes","authors":"Hatef Rahim Sabbaghizadeh , Arshin Oskoueian , Amir Hossein Ashtari","doi":"10.1016/j.repbre.2024.12.001","DOIUrl":"10.1016/j.repbre.2024.12.001","url":null,"abstract":"<div><div>Single-wall carbon nanotubes (SWCNTs) have emerged as promising nanocarriers for targeted cancer drug delivery due to their unique structural properties. However, their cytotoxicity remains a significant challenge, as the biocompatibility of SWCNTs with human cells, particularly hepatocytes, is crucial for their clinical application. The toxicity of SWCNTs is influenced by factors such as nanoparticle size, morphology, surface chemistry, and the presence of impurities. In this study, we aimed to synthesize highly pure SWCNTs and assess their biocompatibility with human hepatocyte cells.</div><div>SWCNTs were fabricated using a modified chemical vapor deposition (CVD) method, followed by a two-step acid purification technique. Raman spectroscopy and electron microscopy confirmed a high purity level of 99.8 %. The biocompatibility of the purified SWCNTs was evaluated using an in vitro model with human hepatocytes. Results indicated that high concentrations of SWCNTs (>50 μg/ml) significantly reduced cell viability, increased lactate dehydrogenase leakage, and elevated lipid peroxidation, while simultaneously suppressing antioxidant enzyme activity.</div><div>Flow cytometry analysis further revealed that exposure to high concentrations of SWCNTs induced apoptosis in hepatocytes. Molecular analysis of key biomarkers demonstrated upregulation of TNF-α, IL1β, NF-kB, and iNOS, alongside downregulation of nrf2 gene and protein expression. These alterations contribute to the mechanisms underlying SWCNT-induced oxidative stress and apoptosis in human hepatocyte cells. Despite the high purity of SWCNTs, their cytotoxic effects may be attributed to their inherent physical properties, including rigidity, surface area, and fiber length.</div><div>In conclusion, while SWCNTs hold great potential for cancer drug delivery, managing their toxicity remains critical for their future therapeutic applications.</div></div>","PeriodicalId":74667,"journal":{"name":"Reproduction and breeding","volume":"5 1","pages":"Pages 29-37"},"PeriodicalIF":0.0,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143175459","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Widening the tool set for breeding superior banana cultivars. Can new techniques of pollen handling and pollination help overcome the lack of recombinant seed in banana breeding? 拓宽香蕉优良品种选育工具。花粉处理和授粉的新技术能否帮助克服香蕉育种中重组种子的缺乏？

Reproduction and breeding

Pub Date : 2024-11-28 DOI: 10.1016/j.repbre.2024.11.001

Ralf Bodo Trognitz

Pollen tube growth (PTG) monitoring was applied to florets of domesticated bananas and plantains. Observations were made in UV microscopy after Aniline Blue staining specific for callose, a carbohydrate of pollen tubes. It was found that total pollen grain numbers on a stigma and total pollen tubes developing after sufficient time for growing to reach the ovules within a female floret ranged from nil to few. This suggested the standard hand pollination technique, HP, may be insufficient to achieve desired pollination quality and respective fertilization for seed development. Pollen deposition and pollen tube growth was at least ten times larger when applying the newly developed pollen-anther-stigma, PAS, technique of pollination. This PAS technique includes mechanically working wilted anther pieces holding and exposing the pollen into the female stigma. PAS provided the potential for maximum seed formation depending on genetic limitations that may be present. PAS is therefore recommended for use in investigations of reproductive processes, such as sterility and self-incompatibility, and it can help increasing the production of recombinant seeds in planned crosses for breeding and selection.

应用花粉管生长（PTG）技术对驯化香蕉和大蕉小花进行了监测。花粉管碳水化合物胼胝质经苯胺蓝染色后在紫外显微镜下观察。在雌花中，柱头上的总花粉粒数和发育到胚珠所需时间的总花粉管数从零到很少不等。这表明标准的手传粉技术HP可能不足以达到理想的授粉质量和种子发育的各自受精。应用新开发的花粉-花药-柱头（PAS）授粉技术，花粉沉积和花粉管生长至少增加10倍。这种PAS技术包括机械加工枯萎的花药片，将花粉暴露在雌性柱头中。PAS根据可能存在的遗传限制提供了最大种子形成的潜力。因此，PAS被推荐用于研究生殖过程，如不育性和自交不亲和，它可以帮助增加重组种子的产量，在计划杂交中进行育种和选择。

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引用次数: 0

Construction of an integrated linkage map of sugarcane using unigene-derived microsatellite (SSR) markers 利用单基因微卫星（SSR）标记构建甘蔗综合联系图

Reproduction and breeding

Pub Date : 2024-11-26 DOI: 10.1016/j.repbre.2024.11.002

Sujeet P. Singh , Jyoti Rasogi , Chethana Ck , R.K. Singh , A. Nigam , Ram Baran Singh

Sugarcane (Saccharum officinarum L.) is one of the key agro-industrial crops that produce raw sugar and bio-fuel ethanol, which contributes to sustainable national economy, food and nutritional security in tropical areas globally. Traditional sugarcane breeding practices are tedious and labour-intensive practice to develop elite high-yielding and resistant cultivars. Sugarcane improvement based on high-throughput techniques to develop traits linked DNA markers through establishing marker-trait associations using linkage analysis. The SSR and markers offer a user-friendly and cost-efficient genomic tool for marker-trait linkage analysis in crop species. In the current study, an integrated sugarcane genetic linkage map was constructed with SSR and TRAP molecular markers using 226 progenies derived from a bi-parental (UP 9530 × Co 86,011) mapping population. The biparental mapping population (266 progenies) was genotyped which generated a total of 815 polymorphic loci with 96 SSR and 11 TRAP primer combinations. Out of 815 markers, 324 markers were scattered onto 107 linkage groups (LGs) and for all the linkage maps, the cumulative genome length was found to be 7608.7 cM, with an average length of 71.11 cM per LG. The length of LGs was ranging from 0.06 to 238.44 cM, with an average of 23.48 cM between two adjacent markers and the number of markers per LG varied from 2 to 11. Out of 324 linked markers, 232 (71.60 %) were found to be single dose (1:1) and 92 (28.39 %) double single dose (3:1) in segregation pattern in genotyping. All the LGs with common markers on individual maps were merged to construct an integrated map using a map integration tool. Future research will be focused on validation of the linked markers in sugarcane involves a combination of phenotypic studies, molecular analysis, genetic mapping, and field trials. This ensures the marker is accurate, reliable, and applicable across different environments and genetic backgrounds. Moreover, the developed SSR-based linkage map will be a useful suit for understanding genetic architecture, marker-assisted breeding for improving yield and quality, identifying disease resistance genes, and accelerating genetic research in polyploid sugarcane and other related polyploidy crop species.

甘蔗（Saccharum officinarum L.）是生产原糖和生物燃料乙醇的主要农用工业作物之一，有助于全球热带地区国民经济的可持续发展以及粮食和营养安全。传统的甘蔗育种方法繁琐且劳动密集，难以培育出高产、抗性强的优良品种。甘蔗改良以高通量技术为基础，通过联系分析建立标记与性状之间的联系，开发与性状相关的 DNA 标记。SSR 和标记为农作物物种的标记-性状关联分析提供了一种用户友好且经济高效的基因组工具。在本研究中，利用双亲（UP 9530 × Co 86,011）制图群体的 226 个后代，使用 SSR 和 TRAP 分子标记构建了甘蔗综合遗传连锁图谱。对双亲制图群体（266 个后代）进行了基因分型，通过 96 个 SSR 和 11 个 TRAP 引物组合共产生了 815 个多态位点。在 815 个标记中，有 324 个标记分布在 107 个连接组（LG）上，所有连接图的基因组累积长度为 7608.7 cM，每个 LG 的平均长度为 71.11 cM。LG 的长度从 0.06 到 238.44 cM 不等，两个相邻标记之间的平均长度为 23.48 cM，每个 LG 的标记数从 2 到 11 不等。在 324 个连锁标记中，232 个（71.60%）为单剂量（1:1），92 个（28.39%）为双单剂量（3:1）。利用图谱整合工具，对单个图谱上具有共同标记的所有 LGs 进行了合并，构建了一个整合图谱。今后的研究重点将是对甘蔗中的连锁标记进行验证，包括表型研究、分子分析、基因图谱绘制和田间试验。这将确保标记准确、可靠，并适用于不同的环境和遗传背景。此外，所开发的基于 SSR 的连接图将成为了解遗传结构、提高产量和品质的标记辅助育种、鉴定抗病基因以及加速多倍体甘蔗和其他相关多倍体作物物种遗传研究的有用工具。

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引用次数: 0

Genetic and cytogenetic analysis of Moringa oleifera short and long capsule phenotypes 油杉长短蒴果表型的遗传和细胞遗传分析

Reproduction and breeding

Pub Date : 2024-11-12 DOI: 10.1016/j.repbre.2024.10.002

Martha Gómez-Martínez , Jesús David García-Ortiz , Susana Gómez-Martínez , Adriana C. Flores-Gallegos , M. Humberto Reyes-Valdés , Jesús A. Morlett-Chávez , Raúl Rodríguez-Herrera

Cytogenetic and molecular studies hold significant importance in plant breeding programs. In the case of moringa, such studies are scarce. Therefore, the objective of this research was to determine the chromosomal number and genetic diversity parameters in two phenotypes of Moringa oleifera Lam. (short and long capsule) using chromosomal analysis and ISSR markers, respectively. Cytogenetic analyses were conducted using the “Somatic chromosomes in root apices” technique, with acetocarmine and papain for staining. It was possible to identify 2N = 28 chromosomes in a single cell of the long capsule phenotype, ranging in size from 0.05 to 0.10 μm. Cells in both moringa phenotypes were observed in prophase, metaphase, anaphase, and telophase. High genetic variability was found in both phenotypes of moringa, as indicated by a Shannon index of 0.81. Additionally, the principal component analysis and UPGMA (Unweighted Pair-Group Method with Arithmetic) groups revealed genetic isolation between the studied moringa phenotypes. The significant polymorphism obtained with most primers suggests that the tested ISSR markers are highly useful for studying genetic diversity in moringa. These findings provide a robust foundation for future research in genetic improvement and conservation of moringa, highlighting the relevance of cytogenetic and molecular studies in this species.

细胞遗传学和分子研究在植物育种计划中具有重要意义。就辣木而言，此类研究很少。因此，本研究的目的是利用染色体分析和 ISSR 标记，确定 Moringa oleifera Lam.(短蒴果和长蒴果）的染色体数目和遗传多样性参数。细胞遗传学分析采用 "根尖体细胞染色体 "技术，并用乙酰卡胺和木瓜蛋白酶进行染色。在长蒴果表型的单个细胞中可以鉴定出 2N = 28 条染色体，大小从 0.05 到 0.10 μm。在原分裂期、移行分裂期、无形变期和端粒期都观察到了这两种形态的细胞。两种表现型的莫林格都具有很高的遗传变异性，香农指数为 0.81。此外，主成分分析和 UPGMA（非加权配对组算术法）分组显示，所研究的辣木表型之间存在遗传隔离。大多数引物都具有明显的多态性，这表明测试的 ISSR 标记对研究辣木的遗传多样性非常有用。这些发现为今后的辣木遗传改良和保护研究奠定了坚实的基础，突出了对该物种进行细胞遗传学和分子研究的意义。

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引用次数: 0

Masculinization? Rare variation in canine length among female water deer (Hydropotes inermis) can lead to errors in sex identification 男性化？雌性水鹿（Hydropotes inermis）犬齿长度的罕见变异可能导致性别鉴定错误

Reproduction and breeding

Pub Date : 2024-11-11 DOI: 10.1016/j.repbre.2024.10.001

Seong-Min Lee

Most cervids feature antlers for mate competition, but water deer possess uniquely elongated maxillary canines instead. This study examined the presence of elongated canines, a trait usually seen in males, in female water deer from 185 individuals culled in South Korea. Measurements of body length, height, and mass were recorded, along with assessments of reproductive capability. The results showed that 1.6 % of the females exhibited elongated canines, retaining their fertility. Their body sizes remained within the typical range for their same age group, with their canines continuing to grow for at least three years. Thus, this rare variation, which is similar to males, may lead to infrequent errors in sex identification based on physical appearance.

大多数鹿科动物都有用于竞争配偶的鹿角，但水鹿却拥有独特的细长上颌犬齿。这项研究考察了在韩国捕杀的 185 头雌性水鹿身上是否存在通常见于雄性的细长犬齿。研究记录了雌鹿的体长、身高和体重，并对其繁殖能力进行了评估。结果显示，1.6% 的雌性水鹿表现出犬齿变长，但仍保持着生育能力。它们的体型保持在同年龄组的典型范围内，犬齿继续生长至少三年。因此，这种罕见的变异与雄性相似，可能会导致根据外貌进行性别鉴定时出现不常见的错误。

引用次数: 0

Semen characteristics, freezability, and application of motility-based protein markers (proAKAP4) in assessing the suitability of superior Bali bulls (Bos sondaicus) at the Regional AI Center 精液特征、可冷冻性和运动性蛋白标记（proAKAP4）在地区人工授精中心评估优质巴厘公牛（Bos sondaicus）适用性中的应用

Reproduction and breeding

Pub Date : 2024-10-15 DOI: 10.1016/j.repbre.2024.09.005

Kudratullah , Raden Iis Arifiantini , Enny Yuliani , Berlin Pandapotan Pardede , Syahruddin Said , Bambang Purwantara

The use of molecular markers in selecting superior bulls, including Bali bulls, for the AI program will significantly increase reproductive efficiency and affect the economic aspects of several countries, especially in the livestock industry sector. Motility-based protein markers, proAKAP4, as one of the molecular markers that have recently been developed in the breeding industry, are expected to be applied and used at AI centers in Indonesia to optimize the use of superior Bali bulls, which are also supported by various semen and freezability characteristics. Using all Bali bulls from the Regional AI center in one of the provinces in Indonesia, this study divided the bulls into motility grade A and motility grade B groups. The collected semen is tested using various methods based on the literature, including fresh and frozen semen and sperm freezability. The proAKAP4 was quantified using the Bull 4MID® Kit with ELISA. The characteristics of fresh semen were significantly different (P < 0.05) except for sperm abnormalities (P > 0.05). The frozen semen characteristics were significantly different (P < 0.05) regarding plasma membrane integrity, DNA fragmentation, and several kinematic parameters. There was no significant difference (P > 0.05) in the freezability characteristics of Bali bull sperm. ProAKAP4 was significantly (P < 0.05) more abundant in the motility grade B bull group and was closely correlated (P < 0.05) with progressive motility and sperm viability. Motility-based protein markers, proAKAP4, can be applied to bulls at the Regional AI Center, with quantification results showing that half of the bull population used in research does not meet the fitness standards for superior bulls.

在人工授精计划中使用分子标记来选择包括巴厘岛公牛在内的优良公牛，将大大提高繁殖效率，并影响多个国家的经济方面，尤其是畜牧业部门。基于运动能力的蛋白标记 proAKAP4 是最近在育种行业开发的分子标记之一，有望在印尼的人工授精中心得到应用和使用，以优化优良巴厘岛公牛的使用，这些公牛还具有各种精液和可冷冻性特征。本研究使用印尼某省地区人工授精中心的所有巴厘公牛，将公牛分为活力 A 级和活力 B 级两组。收集的精液根据文献采用各种方法进行检测，包括新鲜和冷冻精液以及精子冷冻性。使用公牛 4MID® 酶联免疫吸附试剂盒对 proAKAP4 进行定量检测。除精子畸形（P >0.05）外，新鲜精液的特征均有显著差异（P <0.05）。冷冻精液在质膜完整性、DNA碎片和几个运动参数方面有明显差异（P < 0.05）。巴厘公牛精子的冷冻特性无明显差异（P > 0.05）。ProAKAP4 在活力等级 B 公牛组中含量明显更高（P <0.05），并且与精子活力和精子存活率密切相关（P <0.05）。以运动能力为基础的蛋白质标记--proAKAP4--可应用于地区人工授精中心的公牛，量化结果显示，用于研究的公牛群体中有一半不符合优良公牛的体能标准。

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引用次数: 0