本文選題：水牛 + 精液品質(zhì)��； 參考：《廣西大學(xué)》2017年碩士論文

【摘要】：本研究通過對(duì)廣西品改站種公牛站44頭尼里-拉菲和33頭摩拉水牛近年的精液生產(chǎn)數(shù)據(jù)的分析,探討品種、氣溫和年齡對(duì)公牛精液品質(zhì)的影響。采用DNA池PCR的方法篩選尼里-拉菲和摩拉水牛HSP70基因的SNP位點(diǎn),通過HRM技術(shù)進(jìn)行基因分型,研究堿基突變和公牛精液品質(zhì)的相關(guān)性。進(jìn)一步對(duì)摩拉和尼里-拉菲水牛中不同精子活力的精液進(jìn)行頂體酶、酸性磷酸酶、α-葡萄糖苷酶活力測(cè)定和HSP60和HSP70蛋白含量定性分析,研究了 HSP70對(duì)公牛精液品質(zhì)的影響,為研究HSP70在精子中的作用機(jī)制和河流型公水牛的選育提供了一定的理論基礎(chǔ)。試驗(yàn)結(jié)果如下:1、結(jié)果表明,摩拉和尼里-拉菲水牛種公牛的精液在精子活力和采精量方面有顯著性的差異(P0.05),尼里-拉菲水牛的精子活力比摩拉水牛高,其采精量要低于摩拉水牛;在尼里-拉菲水牛中,3-5歲種公牛的精子密度和抗凍性能都要顯著的高于6-9歲和10-16歲組公牛(P0.05),3-5歲的種公牛的精子抗凍活力顯著高于10-16歲的公牛(P0.05),高于6-9歲的公牛但不顯著(P0.05)。摩拉水牛精液品質(zhì)的年齡因素影響分析結(jié)果顯示:3個(gè)年齡段水牛種公牛的精子活力、精子凍后活力和采精量都沒有顯著性的差異,6-9歲的種公牛精子密度和活精子數(shù)要顯著高于10-16歲組(P0.05)。從精子抗凍性方面看,10-16歲組的公牛是最低的,只有0.44,3-5歲的公牛最高,達(dá)到了 0.47,有顯著性的差異(P0.05);莫拉和尼里-拉菲水牛的精液品質(zhì)在冬季(1-3月)是最差的,精子活力顯著低于夏季(6-10月)和春秋季(4-5 月、11-12 月)(P0.05)。2、在HSP70基因CDS區(qū)檢測(cè)到3個(gè)SNP位點(diǎn),分別是CDs區(qū)602bp位點(diǎn)、708bp和1284bp位點(diǎn),其中602位點(diǎn)為有義突變,導(dǎo)致了甘氨酸→谷氨酸突變,另2處為無義突變。統(tǒng)計(jì)分析結(jié)果表明:602位點(diǎn)的不同基因型顯著影響了摩拉和尼里-拉菲水牛的精子活力,活精子數(shù)以及精子抗凍能力(P0.05),但對(duì)公水牛采精量和精子密度沒有顯著性的影響,708位點(diǎn)和1284位點(diǎn)對(duì)公牛精液品質(zhì)無顯著性影響(P0.05)。3、在不同活力的摩拉和尼里-拉菲水牛精液的頂體酶、酸性磷酸酶、α-葡萄糖苷酶三種酶活力測(cè)定中,高活力組的頂體酶和α-葡萄糖苷酶顯著高于低活力組的精液(P0.05),但酸性磷酸酶與精子活力相關(guān)性不顯著(P0.05)。4、通過HSP60和HSP70蛋白的Western blotting分析,發(fā)現(xiàn)與低活力組相比,高活力組的HSP70蛋白要顯著高于低活力組(P0.05),其HSP60蛋白含量無顯著性差異(P0.05)。
[Abstract]:In this study, the semen production data of 44 Niri-Lafite and 33 Mora buffaloes were analyzed to explore the effects of breed, temperature and age on the semen quality of bulls. The SNP loci of HSP70 gene of Niri-Lafite and Mora buffalo were screened by DNA pool PCR, and the relationship between base mutation and bull semen quality was studied by HRM technique. Furthermore, acrosome enzyme, acid phosphatase, 偽 -glucosidase activity and HSP60 and HSP70 protein content in semen of different sperm motility of Mora and Nili-Lafite buffalo were determined, and the effect of HSP70 on the semen quality of bull was studied. It provides a theoretical basis for the study of the mechanism of HSP70 in spermatozoa and the breeding of river male buffalo. The results were as follows: 1. The results showed that there was a significant difference in sperm motility and sperm yield between Mora and Near-Lafite buffalo, and the sperm motility of Neri Lafite Buffalo was higher than that of Moray Buffalo. The amount of semen collected was lower than that of Mora buffalo. The spermatozoa density and antifreeze activity of 3-5-year-old bull were significantly higher than those of 6-9 year old and 10-16 year old group in Nili Lafite buffalo. The spermatozoa antifreeze activity of breed bull was significantly higher than that of 10-16 years old and 6-9 years old, but it was not significantly different from that of 6-9 years old and 10-16 years old. The age factors influencing semen quality of buffalo showed that the sperm motility of three age buffalo bulls. There was no significant difference in spermatozoa motility and spermatozoa collection after freezing. The sperm density and sperm count of the bull aged 6-9 years were significantly higher than that of the 10-16 years old group (P0.05). From the point of view of sperm antifreeze, the bull of 10-16 years old group was the lowest, and only 0.445-year-old bull had the highest value (0.47, P 0.05), and the semen quality of Mora and Nili-Lafite buffalo was the worst in the winter (from January to March). The motility of spermatozoa was significantly lower than that in summer (June to October) and in spring and autumn (April to December). Three SNP loci were detected in the CDS region of HSP70 gene, namely CDs 602bp locus 708bp and 1284bp locus. Among them, 602 locus was a seminal mutation, which led to glycine glutamate mutation. The other two were nonsense mutation. The results of statistical analysis showed that different genotypes at the 1: 602 locus significantly affected the sperm motility of Mora and Nili Lafite buffalo. The number of live spermatozoa and the antifreeze ability of spermatozoa were P0.05, but there was no significant effect on the amount of spermatozoa and sperm density in male buffalo. There was no significant effect on the semen quality of bull at locus 708 and locus 1284. There was no significant effect on the semen quality of bull. There was no significant effect on the semen quality of bull at different motility of Morrah and Nili-Lafite. The acrosome enzyme of bovine semen, The activity of acid phosphatase and 偽 -glucosidase, The levels of acrosin and 偽 -glucosidase in high activity group were significantly higher than those in low activity group, but the correlation between acid phosphatase and sperm motility was not significant. Western blotting analysis of HSP60 and HSP70 protein showed that compared with low activity group, there was no significant correlation between ACP and sperm motility. The HSP70 protein in the high activity group was significantly higher than that in the low activity group, and the content of HSP60 protein in the high activity group was not significantly different from that in the low activity group.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S823.83

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