本文關(guān)鍵詞： 中國荷斯坦奶牛 熱應(yīng)激 IL8基因 SNP　出處：《南京農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：我國大部分地區(qū)夏季持續(xù)出現(xiàn)高溫天氣,熱應(yīng)激已成為目前奶牛養(yǎng)殖業(yè)亟待解決的問題之一。奶牛在熱應(yīng)激情況下,機(jī)體新陳代謝發(fā)生紊亂、生產(chǎn)性能及繁殖性能下降、免疫力減弱,更容易遭受乳房炎、子宮內(nèi)膜炎等疾病困擾;同時(shí)還會(huì)造成懷孕母牛流產(chǎn)率、死胎率增加,給養(yǎng)殖場(chǎng)帶來巨大的經(jīng)濟(jì)損失。因此,開展與奶牛抗應(yīng)激相關(guān)基因遺傳多態(tài)性與耐熱性的關(guān)聯(lián)分析,可以為耐熱高產(chǎn)優(yōu)質(zhì)奶牛的分子標(biāo)記輔助育種提供理論依據(jù)和技術(shù)支持。本試驗(yàn)從浙江省杭州蕭山富倫奶牛場(chǎng)選取112頭處于熱應(yīng)激時(shí)期的泌乳中期中國荷斯坦奶牛,運(yùn)用PCR-SSCP法篩查ISG15和IL8基因內(nèi)的SNPs,并進(jìn)行了基因SNPs與個(gè)體直腸溫度、血常規(guī)和血液生化指標(biāo)等的關(guān)聯(lián)分析,結(jié)果如下:(1)IS15基因中檢測(cè)到1個(gè)中度多態(tài)SNP,IL8基因內(nèi)檢測(cè)到11個(gè)中度多態(tài)SNPs(0.25PIC0.5),其中 IL8 基因 g.2209T→G、g.2222G→A、g.2373 G→A 顯著偏離Hardy-Weinberg平衡狀態(tài)。(2)ISG15基因g.844G→A與中國荷斯坦奶牛輕度熱應(yīng)激(THI=76.1)時(shí)的直腸溫度、血常規(guī)和血液生化指標(biāo)不存在顯著關(guān)聯(lián)。(3)IL基因 g.-110G→A、g.1454G→T、g.2373 G→A、g.2784A→G、g.2857 T→C、g.2983G→A與平均血小板體積(MPV)存在顯著關(guān)聯(lián)(P0.05),其中g(shù).2784 A→G、g.2857 T→C、g.2983 G→A位點(diǎn)突變可顯著影響谷草轉(zhuǎn)氨酶(AST)活力(P0.05),突變純合型(GGCCAA)個(gè)體AST活力顯著低于雜合型(AGTCGA)個(gè)體(P0.05),極顯著低于野生純合型(AATTGG)個(gè)體(P0.01),呈現(xiàn)出GGCCAAAGTCGAAATTGG的趨勢(shì),且GGCCAA型個(gè)體血清中鉀離子(K+)含量也處于最低水平。IL8基因g.202C→T、g.264C→T、g.302A→G可顯著影響紅細(xì)胞計(jì)數(shù)(RBC)、紅細(xì)胞壓積(HCT)、血小板計(jì)數(shù)(PLT)、血紅蛋白(HGB)與丙二醛(MDA)含量,此位點(diǎn)突變純合型(TTTTGG)個(gè)體MAD含量顯著低于雜合型(CTCTAG)個(gè)體(P0.05),呈現(xiàn)出TTTTGGCCCCAACTCTAG的趨勢(shì),且TTTTGG型個(gè)體血清中K+含量和直腸溫度較其他基因型處于較低水平。IL8 I基因的 g.202C→T、g.264C→T、g.302A→G 與 g.202 C→T、g.264C→T、g.302 A→G位點(diǎn)可以作為奶牛耐熱性狀候選功能性SNPs,用于耐熱高產(chǎn)奶牛的分子標(biāo)記輔助育種。
[Abstract]:Heat stress has become one of the problems to be solved in dairy cattle breeding industry. Under the condition of heat stress, the metabolism of dairy cattle is disturbed, and the performance of production and reproduction is decreased. Weakened immunity is more likely to suffer from diseases such as mastitis, endometritis, and other diseases. At the same time, it will also result in the abortion rate of pregnant cows and an increase in the stillbirth rate, which will bring huge economic losses to the farms. To analyze the association between genetic polymorphism and heat tolerance of anti-stress genes in dairy cattle. It can provide theoretical basis and technical support for molecular marker-assisted breeding of heat-resistant, high-yield and high-quality dairy cows. 112 Chinese Holstein cows in the middle lactation period of heat stress period were selected from Fulun dairy farm in Xiaoshan, Zhejiang Province. SNPs in ISG15 and IL8 genes were screened by PCR-SSCP method, and the correlation between SNPs gene and rectal temperature, blood routine examination and blood biochemical indexes were analyzed. The results were as follows: 1 moderately polymorphic SNPl8 gene was detected in the IS15 gene, and 11 moderately polymorphic SNPs were detected in 0.25PIC0.5, among which the IL8 gene g.2209T was detected. 鈫扜,g.2222G. 鈫扐,g.2373 G. 鈫扐 significantly deviated from the Hardy-Weinberg equilibrium. G844G of the ISG15 gene. 鈫扵here was no significant correlation between rectal temperature, blood routine examination and blood biochemical index in Chinese Holstein cows with mild heat stress (THIHI-76.1). 鈫扐,g.1454G. 鈫扵,g.2373 G. 鈫扐,g.2784A. 鈫扜,g.2857 T. 鈫扖,g.2983G. 鈫扵here was a significant correlation between P0.05A and mean platelet volume (MPV), among which g. 2784 A. 鈫扜,g.2857 T. 鈫扖,g.2983 G. 鈫扵he mutation of A locus significantly affected the activity of aspartate (P0.05), and the activity of AST in individuals with GGCCAA was significantly lower than that in individuals with heterozygote AGTCGA, and was significantly lower than that in individuals with wild homozygous type (P0.01), showing the trend of GGCCAAAGTCGAAATTGG. The level of K + K in serum of individuals with GGCCAA type is also at the lowest level. IL8 gene g.202C. 鈫扵,g.264C. 鈫扵,g.302A. 鈫扜 could significantly affect the contents of RBC, HCT, PLT, HGB and malondialdehyde (MDA). The MAD content of homozygous individuals was significantly lower than that of heterozygous CTCTAG (P0.05), showing the trend of TTTTGGCCCCAACTCTAG. And the serum K content and rectal temperature of TTTTGG individuals were lower than those of other genotypes. IL8I gene was at the level of g.202C. 鈫扵,g.264C. 鈫扵,g.302A. 鈫扜 and g.202 C. 鈫扵,g.264C. 鈫扵,g.302 A. 鈫扜 locus can be used as functional SNPs for heat tolerance traits of dairy cattle, and it can be used in molecular marker assisted breeding of heat tolerance and high yield cows.
【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S823

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