【摘要】：本實驗選用糯性青稞品種甘墾5號和非糯性青稞品種北青6號和昆侖12號作為試驗材料,以雙波長分光光度法測定3個青稞品種籽粒灌漿期中直鏈淀粉與支鏈淀粉含量的動態(tài)變化;通過測定酶活分析研究了淀粉代謝的關鍵酶活性變化;通過RT-PCR研究并分析了淀粉代謝關鍵酶編碼基因的表達量,以及不同基因表達量之間和基因表達量與對應酶活的相互關系。結果表明:1.隨著籽粒灌漿期的進行甘墾5號、北青6號和昆侖12號的支鏈淀粉的含量、總淀粉的含量隨籽粒灌漿期進行而逐漸增加,北青6號和昆侖12號的直鏈淀粉的含量也隨之逐漸增加。其中北青6號和昆侖12號的支鏈淀粉前期積累速率較快,而直鏈淀粉后期積累速率較快,最終導致支鏈淀粉和直鏈淀粉的比例逐漸下降,花后15天左右,其比例已無差異。2.甘墾5號、北青6號和昆侖12號籽粒灌漿過程中催化淀粉合成的關鍵酶:淀粉合成酶AGPP、SSS、GBSS和SBE的活性變化均呈單峰曲線,北青6號的AGPP和昆侖12號的GBSS的活性在花后25天左右達到峰值,而甘墾5號的AGPP、SSS和SBE的活性和北青6號的SSS、GBSS和SBE的活性以及昆侖12號的AGPP、SSS、和SBE均在花后20天左右達到峰值;而在籽粒灌漿過程中α-淀粉酶則出現(xiàn)升-降-升-降的變化趨勢,即在花后10天左右出現(xiàn)第一個峰值,花后20天左右降到最低,花后25天左右,再次出現(xiàn)峰值,之后開始下降,而β-淀粉酶基本上是呈先下降后上升的趨勢,在花后20天左右活性值最低。3.相關性分析結果表明,北青6號籽粒灌漿期淀粉合成酶AGPP、SSS、GBSS和SBE的活性變化與直鏈淀粉含量的變化呈顯著正相關,北青6號和昆侖12號籽粒灌漿直鏈淀粉、支鏈淀粉和總淀粉積累速率與AGPP、GBSS、SSS和SBE活性變化顯著或極顯著正相關。在甘墾5號中未檢測到直鏈淀粉的積累,甘墾5號籽粒灌漿期的支鏈淀粉和總淀粉積累速率與AGPP和SSS呈顯著或者極顯著正相關。而北青6號和昆侖12號籽粒灌漿期的直鏈淀粉、支鏈淀粉和總淀粉的積累速率與β-淀粉酶活性呈顯著或極顯著負相關。甘墾5號籽粒灌漿期的支鏈淀粉和總淀粉積累速率與α-淀粉酶呈顯著負相關。青稞籽粒灌漿過程中淀粉含量積累是由于淀粉合成酶活性的增強,而不是分解酶活性的減弱。因此,青稞籽粒中淀粉積累是淀粉代謝酶(合成和分解方向)綜合作用的結果。4.基因的表達量分析結果表明:甘墾5號、北青6號和昆侖12號的淀粉合成編碼基因的表達模式大致可分為兩種曲線模式,即低-高-低的單峰曲線和高-低-高-低的雙峰曲線。其中GBSSI在糯性青稞品種甘墾5號中一直保持低量表達,但在非糯性材料北青6號和昆侖12號中呈現(xiàn)單峰曲線的表達模式;AGPP、GBSSI、SSSIII和SBEI在胚乳淀粉合成時期表達量較高;GBSSII后期表達量較高,但在整個灌漿過程中表達量均較低。淀粉分解編碼基因中,AMY4的mRNA在3個青稞品種整個開花過程中呈現(xiàn)逐漸升高的趨勢外,AMY1、AMY2和AMY3均呈現(xiàn)在開花前期表達量較高,開花中期表達量相對較低但保持穩(wěn)定,開花后期表達量逐漸回升的趨勢。青稞籽粒灌漿過程中BAM1和BAM2在開花前期表達量較高,而BAM6和BAM7則在開花后期有較高的表達量,BAM4和BAM5在整個灌漿過程中則為恒量表達�？赏ㄟ^分解酶基因表達量解釋淀粉分解酶活性先降低后上升的變化趨勢。
[Abstract]:In this experiment, glutinous and non-glutinous highland barley varieties Ganken 5 and Kunlun 12 were selected as experimental materials, and the dynamic changes of amylose and amylopectin contents in grain filling stage of three highland barley varieties were determined by dual-wavelength spectrophotometry. The expression of genes encoding key enzymes in starch metabolism was studied by RT-PCR, and the relationship between gene expression and corresponding enzyme activity was analyzed. The results showed that: 1. With the content of amylopectin in Ganken 5, Beiqing 6 and Kunlun 12 during grain filling, the total starch content increased with grain filling. The amylopectin content of Beiqing 6 and Kunlun 12 increased gradually, and the amylopectin accumulation rate of Beiqing 6 and Kunlun 12 was faster in the early stage, but the amylopectin accumulation rate was faster in the later stage, which eventually led to the gradual decrease of the amylopectin and amylopectin proportion, and the proportion had no difference about 15 days after anthesis. 2. The activities of the key enzymes for starch synthesis during grain filling: starch synthase AGPP, SSS, GBSS and SBE showed a single peak curve. The activities of AGPP and GBSS of Beiqing 6 and Kunlun 12 reached a peak value about 25 days after anthesis, while those of AGPP, SSS and SBE of Ganken 5 and SSS, GB of Beiqing 6 reached a peak value. The activity of SS and SBE, AGPP, SSS, and SBE of Kunlun 12 reached their peak value about 20 days after anthesis, while the change trend of alpha-amylase appeared during grain filling, that is, the first peak value appeared about 10 days after anthesis, the lowest value appeared about 20 days after anthesis, the peak value appeared again about 25 days after anthesis, and then began to decline. The activity of amylase AGPP, SSS, GBSS and SBE in grain filling stage of Beiqing 6 was positively correlated with amylose content. The amylose content in grain filling stage of Beiqing 6 and Kunlun 12 was positively correlated with amylose content. Amylose and total starch accumulation rates were significantly or extremely significantly positively correlated with AGPP, GBSS, SSS and SBE activities. Amylose accumulation was not detected in Ganken 5. Amylose and total starch accumulation rates at grain filling stage of Ganken 5 were significantly or very significantly positively correlated with AGPP and SSS. The accumulation rates of amylopectin, amylopectin and total starch were significantly or extremely significantly negatively correlated with the activity of beta-amylase. The accumulation rates of amylopectin and total starch were significantly negatively correlated with the activity of alpha-amylase at grain filling stage of Ganken 5. The accumulation of starch content during grain filling was due to the increase of amylase synthase activity, not to the increase of amylase synthase activity. Therefore, starch accumulation in highland barley grains is the result of the synthesis and decomposition of starch metabolizing enzymes. 4. The results of gene expression analysis show that the expression patterns of starch synthesis coding genes in Ganken 5, Beiqing 6 and Kunlun 12 can be divided into two kinds of curve patterns, that is, low-high-low single peak curve. The expression of GBSSI in waxy highland barley variety Ganken 5 was low, but in non-waxy materials Beiqing 6 and Kunlun 12, the expression of AGPP, GBSSI, SSSIII and SBEI was high at the endosperm starch synthesis stage; the expression of GBSSII was high at the late stage, but in the whole irrigation stage. The expression of AMY4 mRNA increased gradually in the whole flowering process of three highland barley varieties. The expression of AMY1, AMY2 and AMY3 were higher in the early flowering stage, lower but stable in the middle flowering stage, and gradually increased in the late flowering stage. During grain filling, BAM1 and BAM2 were highly expressed in the early flowering stage, while BAM6 and BAM7 were highly expressed in the late flowering stage, while BAM4 and BAM5 were constant in the whole filling stage.
【學位授予單位】：石河子大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S512.3

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相關期刊論文 前10條

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本文編號：2240352