本文選題：紅羅非魚 + 耐寒性��； 參考：《南京農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：體色是魚類一種較獨特的表型性狀,有重要的生物多樣性價值和遺傳學(xué)理論研究價值。魚類體色變化是一個復(fù)雜的過程,遺傳因素以及水溫、光照、pH值、溶氧等眾多外界環(huán)境因素都會造成魚類體色的變化。在紅羅非越冬養(yǎng)殖期間發(fā)現(xiàn),經(jīng)歷過低溫的紅羅非魚通常會發(fā)生體色變化,出現(xiàn)紅色或者黑色斑紋,這樣的變化會使紅羅非魚的經(jīng)濟價值下降。紅羅非魚是暖水性魚類,具有不耐低溫的特性。推斷低溫會影響紅羅非魚體色變化。本文從血清生理生化指標(biāo)、抗氧化酶及兩個抗寒候選基因角度分析比較了以色列、臺灣和馬來西亞三個紅羅非魚品系的耐寒性差異。為了進一步深入探討紅羅非魚體色變化的機制,從黑色素合成通路的分子角度比較了6個體色相關(guān)基因(MC1R、TYRP1、ASIP、DCT、SOX10和S1C7A11)在三個紅羅非魚品系中的表達差異。主要研究結(jié)果如下:1.三個不同品系紅羅非魚耐寒性能比較實驗選取平均體重為500g左右的以色列品系、臺灣品系和馬來西亞品系的紅羅非魚作為實驗對象,設(shè)定19℃的常溫對照組和從19℃開始每天降低1℃直到8℃的冷應(yīng)激實驗組,每組設(shè)定2個平行組,每個平行組中6尾魚。對照組在實驗開始時進行采樣,實驗組在魚開始出現(xiàn)死亡情況時進行采樣。測定血清丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST)、乳酸脫氫酶(LDH)、血糖(GLU)、總蛋白(TP)、K+、Na+、Cl-、總超氧化物歧化酶、丙二醛和超微量Na+-K+-ATP酶。結(jié)果表明:以色列和臺灣品系在10.3℃開始出現(xiàn)死亡個體,馬來西亞品系的耐寒性能更強一點,在9.3℃出現(xiàn)死亡個體。三個品系紅羅非魚在8.3℃全部死亡。羅非魚可在6-10℃低溫下短暫存活,但是無法存活24h以上。以色列和臺灣品系死亡溫度比馬來西亞品系高1℃。與對照組相比,三個品系的實驗組ALT、AST、LDH、GLU含量顯著上升(P0.05),以色列和臺灣品系TP含量顯著性下降(P0.05),而馬來西亞品系TP含量顯著上升(P0.05)。以色列品系SOD含量顯著上升(P0.05),MDA含量顯著上升(P0.05),Na+-K+-ATP酶含量顯著下降(P0.05);臺灣品系和馬來西亞品系SOD含量顯著下降(P0.05),Na+-K+-ATP酶含量顯著上升(P0.05)。三種品系的耐寒性能不同,所以體內(nèi)的抗氧化酶含量以及活性也并不相同,其中馬來西亞品系經(jīng)歷的溫度更低,魚體內(nèi)臟等受損程度更加嚴(yán)重,所以體內(nèi)抗氧化酶等不能發(fā)揮其免疫的作用,而是三種品系抗氧化酶等活性表達量不同。當(dāng)受到低溫脅迫時,SCD基因在以色列品系、臺灣品系肝臟中mRNA表達量顯著上升,SCD基因在馬來西亞品系肝臟中mRNA表達量顯著下降(P0.05)。HSP70基因在以色列品系、臺灣品系和馬來西亞品系肝臟中mRNA表達量顯著上升(P0.05)�？梢�,急性低溫應(yīng)激可以引起紅羅非魚體的生理機能的變化,其中馬來西亞品系紅羅非的耐寒性比以色列和臺灣品系紅羅非耐寒性能可能更強。2、以色列、臺灣、馬來西亞品系紅羅非體色相關(guān)基因的表達分析(1)馬來西亞紅羅非6個體色相關(guān)基因的表達分析實驗材料選擇身體健壯,健康狀況良好的馬來西亞紅羅非“紅斑”、“粉白”、“黑斑”三種體色類型的各3尾。每尾魚采集背部皮膚、腹部皮膚、肌肉、腸、心、腦共六種組織。qRT-PCR 分析表明,MC1R、ASIP、TYRP1、DCT 和 SLC7A11基因在皮膚組織中表達量顯著高于其他四個組織中的表達量(P0.05),在心臟和腸中表達量次之,在肌肉和腦中表達量最低;SOX10基因在腦中表達量顯著高于其他四個組織(P0.05),在皮膚及肌肉中表達量次之,在腸和心中表達量最低。MC1R、ASIP、TYRP1、SOX10和SLC7A11基因在紅斑體色類型皮膚中的表達量顯著地高于其他體色(P0.05)。DCT基因在粉白體色類型和黑斑體色類型的皮膚組織中表達量顯著的高于紅斑體色類型(P0.05)。由此可以得知體色類型并不是由一個基因控制的,是由一個復(fù)雜的基因網(wǎng)絡(luò)進行調(diào)控,各基因之間相互作用最終決定與體色類型。(2)三個品系紅羅非皮膚組織中6個基因的表達差異分析實驗材料選擇身體健壯,健康狀況良好的紅羅非魚。馬來西亞、臺灣和以色列品系各取3尾的背部和腹部皮膚組織。對這三種紅羅非品系的背部和腹部皮膚組織進行了基因mRNA的表達差異比較。qRT-PCR分析表明,在背部皮膚組織中,MC1R、ASIP、TYRP1、DCT、SOX10和SLC7A11基因在馬來西亞品系表達量顯著的高于其他兩個品系(P0.05),ASIP和TYRP1基因在以色列品系中的表達量顯著的高于臺灣品系中的表達量(P0.05),MC1R、DCT、SOX10和SLC7A11基因在以色列和臺灣品系中的表達量沒有顯著性差異(P0.05)。在腹部皮膚組織中,MC1R、ASIP、DCT和SOX10基因在馬來西亞品系表達量最高,TYRP1和SLC7A11基因在臺灣品系中表達量最高。TYRP1和SLC7 A1 1基因在以色列、臺灣與馬來西亞品系中均有顯著性差異(P0.05),MC1R、ASIP和SOX10基因在以色列與臺灣品系中均存在顯著性差異(P0.05)。由此可以得知體色相關(guān)基因的表達量與魚種類有一定的相關(guān)性。(3)6個基因在各品系紅羅非魚背腹部皮膚組織中表達差異馬來西亞、臺灣和以色列品系各取3尾的背部和腹部皮膚組織。對每種品系紅羅非品系的背部和腹部皮膚組織中的6個基因mRNA的表達差異進行比較。qRT-PCR分析表明,在以色列品系中,MC1R、TYRP1、DCT、SOX10和SLC7A11基因在背部皮膚組織的表達量顯著的高于腹部皮膚(P0.05);在臺灣品系中,ASIP基因在腹部皮膚組織中表達量顯著的高于背部皮膚(P0.05),MC1R、DCT、SOX10和SLC7A11基因在背部的表達量顯著高于腹部皮膚(P0.05)。在馬來西亞品系中,MC1R、ASIP、TYRP1、SOX10和SLC7A11基因在背部的表達量顯著高于腹部皮膚(P0.05)。由此可以得知,ASIP基因在紅羅非魚的背部和腹部皮膚組織表達有差異。
[Abstract]:Body color is a more unique phenotypic character of fish. It has important value of biological diversity and genetic theory. The change of fish body color is a complex process. Many external environmental factors such as genetic factors and water temperature, light, pH value, dissolved oxygen and other environmental factors will cause changes in the body color of fish. Red tilapia usually changes color and appears red or black markings. This change will reduce the economic value of red tilapia. Red tilapia is warm water fish and has the characteristics of low temperature resistance. It is inferred that low temperature will affect the body color change of red tilapia. This article from serum physiological and biochemical indexes, antioxidant enzymes and two The cold tolerance difference between three red tilapia lines in Israel, Taiwan and Malaysia was analyzed by an angle of cold resistant candidate genes. In order to further explore the mechanism of the body color change of the red tilapia, 6 color related genes (MC1R, TYRP1, ASIP, DCT, SOX10 and S1C7A11) were compared in three red ROS from the molecular angle of the melanin synthesis pathway. The main results are as follows: 1. the comparison experiment of the cold resistance of three different strains of red tilapia selected the Israeli strains with the average weight of about 500g, the Taiwan strain and the red tilapia of Malaysia strain as the experimental object, setting the normal temperature control group at 19 degrees C and decreasing 1 degrees to 8 every day from 19 degrees C. In the cold stress experimental group, 2 parallel groups were set in each group, 6 fish in each parallel group. The control group was sampled at the beginning of the experiment. The experimental group was sampled at the beginning of the death of the fish. The serum prop aminotransferase (ALT), AST, LDH, GLU, TP, K+, Na+, Cl-, and total superoxide dismutase were measured. Enzyme, malondialdehyde and ultramicro Na+-K+-ATP enzyme. The results showed that the death individual began to appear in Israel and Taiwan strain at 10.3 degrees C, the cold resistance of Malaysia strain was stronger and the death individual appeared at 9.3. Three strains of red tilapia died at 8.3. The tilapia could survive at 6-10 temperature, but could not survive 24h The death temperature of Israel and Taiwan strains was 1 higher than that of the Malaysia strain. Compared with the control group, the content of ALT, AST, LDH, GLU in the experimental group increased significantly (P0.05), the TP content in Israel and Taiwan decreased significantly (P0.05), while the TP content in the Malaysia strain increased significantly (P0.05). The SOD content of the Israeli strain rose significantly (P0). .05), the content of MDA increased significantly (P0.05), and the content of Na+-K+-ATP enzyme decreased significantly (P0.05); the content of SOD in Taiwan strain and Malaysia strain decreased significantly (P0.05), and the content of Na+-K+-ATP enzyme increased significantly (P0.05). The cold resistance of the three strains was different, so the content and activity of antioxidant enzymes in the body were different, among which the Malaysia strain was experienced. The temperature is lower and the damage degree of the fish viscera is more serious, so the antioxidant enzyme in the body can not play its immune function, but the active expression of antioxidant enzymes in the three strains of the three strains is different. When under low temperature stress, the expression of mRNA in the liver of the Taiwan strain increases significantly in the Israeli strain, and the SCD gene is in the Malaysia strain. The expression of mRNA in the liver decreased significantly (P0.05).HSP70 gene in the Israeli strains, the Taiwan strain and the Malaysia strain liver mRNA expression increased significantly (P0.05). The acute low temperature stress could cause the physiological function changes of the red tilapia, and the cold resistance of the Malaysia strain red rose was more red than that of the Israeli and Taiwan strains. The cold resistance of Luo Fei may be more.2, Israel, Taiwan, Malaysia and Malaysia, the expression analysis of non body color related genes (1) the expression analysis of 6 color related genes in red Rome, Malaysia, three kinds of body color, red spot, pink and black spot in Malaysia, healthy and healthy The.QRT-PCR analysis of six tissues, including back skin, abdominal skin, muscle, intestine, heart and brain, showed that the expression of MC1R, ASIP, TYRP1, DCT and SLC7A11 genes in the skin tissues was significantly higher than those of the other four tissues (P0.05), and the lowest expression in the heart and intestines, and the lowest expression in the muscles and brain; SOX1. The expression of 0 gene in the brain was significantly higher than that of the other four tissues (P0.05), the expression in the skin and muscle was the lowest, and the expression in the intestine and heart was lowest.MC1R. The expression of ASIP, TYRP1, SOX10 and SLC7A11 genes in the erythematous color type skin was significantly higher than that of the other body color (P0.05).DCT genes in the pink and black body color types. The expression of the skin tissue is significantly higher than that of the erythema color type (P0.05). Therefore, it can be found that the body color type is not controlled by a gene, and is regulated by a complex gene network. The interaction of each gene is ultimately determined by the type of body color. (2) the difference of the expression of 6 genes in the three strains of red non skin tissue is divided. The experimental materials selected the healthy and healthy red tilapia, healthy red tilapia, Malaysia, Taiwan, and the Israeli Department of Malaysia, each took the back and abdominal skin tissues of the 3 tails. The difference of gene expression between the back and abdominal skin tissues of the three red non strain lines was compared with.QRT-PCR analysis, in the back skin tissue, MC1R, ASIP, The expression of TYRP1, DCT, SOX10 and SLC7A11 genes in Malaysia strain was significantly higher than that of other two lines (P0.05). The expression of ASIP and TYRP1 genes in Israeli strains was significantly higher than that in Taiwan strain (P0.05), and there was no significant difference in the expression of MC1R, DCT, SOX10 and SLC7A11 genes in Israel and Taiwan strains. .05). In the abdominal skin tissue, the highest expression of MC1R, ASIP, DCT and SOX10 genes in Malaysia strain, the highest.TYRP1 and SLC7 A1 1 genes of TYRP1 and SLC7A11 genes in Taiwan, Taiwan and Malaysia are significant differences (P0.05). There was a significant difference (P0.05). It was found that the expression of body color related genes was related to the fish species. (3) the 6 genes expressed in the dorsal and abdominal skin tissues of the strains of red tilapia, Malaysia, Taiwan and Israel, each took 3 back and abdominal skin tissues. The difference in expression of 6 genes mRNA in the back and abdominal skin tissues was compared with.QRT-PCR analysis. In the Israeli strain, the expression of MC1R, TYRP1, DCT, SOX10 and SLC7A11 genes in the back skin tissues was significantly higher than that of the abdominal skin (P0.05); in the Taiwan strain, the expression of the ASIP gene in the abdominal skin tissues was significantly higher than that in the abdominal skin. The back skin (P0.05), MC1R, DCT, SOX10 and SLC7A11 genes were significantly higher in the back than in the abdominal skin (P0.05). In the Malaysia strain, the expression of MC1R, ASIP, TYRP1, SOX10 and SLC7A11 genes in the back was significantly higher than that of the abdominal skin (P0.05). There is a difference.
【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S917.4

【參考文獻】

相關(guān)期刊論文 前10條

1 張建一;梁春年;吳曉云;張良斌;郭憲;裴杰;閻萍;;牦牛Agouti基因的克隆及編碼區(qū)多態(tài)性研究[J];華北農(nóng)學(xué)報;2014年05期

2 楊竹青;王自蕊;熊六鳳;阮記明;隗黎麗;;MC1R基因在中國草金魚中的遺傳變異及其進化分析[J];江西農(nóng)業(yè)大學(xué)學(xué)報;2014年05期

3 朱艷菲;胡義潔;吳明明;李禎;孫金海;張廷榮;;紅褐色烏蘇里貉TYRP1基因的克隆及表達水平分析[J];中國畜牧雜志;2014年11期

4 董晶晶;謝駿;習(xí)丙文;;魚類黑素皮質(zhì)素受體(MCR)研究進展[J];中國水產(chǎn)科學(xué);2014年03期

5 楊民紅;李蘭會;;動物皮毛色澤形成機制研究進展[J];貴州畜牧獸醫(yī);2013年05期

6 李洪濤;曾獻存;張文祥;趙海璇;惠文巧;劉鋼;羅燕;班謙;趙宗勝;賈斌;;哈薩克綿羊MC1R和ASIP基因多態(tài)性及表達量與被毛顏色表型相關(guān)性的研究[J];畜牧獸醫(yī)學(xué)報;2013年03期

7 常玉梅;徐麗華;李世國;唐然;梁利群;;鯉耐低溫候選基因CcSCD全長cDNA的克隆及功能預(yù)測[J];水產(chǎn)學(xué)雜志;2012年03期

8 趙彥斌;孫兆增;白杰英;張小飛;李愛學(xué);柳巨雄;隋麗華;胡仲明;曾林;;黑線倉鼠及其白化突變系TYR、TYRP1的基因表達水平比較分析[J];中國比較醫(yī)學(xué)雜志;2012年07期

9 ;Production of Brown/Yellow Patches in the SLC7A11 Transgenic Sheep via Testicular Injection of Transgene[J];遺傳學(xué)報;2012年06期

10 劉波;王美W，

本文編號：2077938