本文選題：玉米 + 絲氨酸/蘇氨酸蛋白磷酸酶��； 參考：《河南農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：絲氨酸/蘇氨酸蛋白磷酸酶6亞基(catalytic subunits of Ser/Thr protein phosphatase 6,PP6C)是PP6全酶的催化亞基。在模式植物擬南芥中的研究表明,PP6C參與生長素極性運(yùn)輸、脫落酸信號(hào)轉(zhuǎn)導(dǎo)和光信號(hào)轉(zhuǎn)導(dǎo)途徑介導(dǎo)的開花調(diào)控。本研究采用RT-PCR方法克隆了ZmPP6C基因,分析了蛋白結(jié)構(gòu)特征與同源蛋白間的進(jìn)化關(guān)系,利用定量PCR的方法分析了ZmPP6C基因?qū)Σ煌赓|(zhì)、黑暗到各種光質(zhì)轉(zhuǎn)換、長日照和短日照處理,以及脅迫處理下的表達(dá)模式,結(jié)果如下:1.蛋白磷酸酶6亞基的氨基酸序列在進(jìn)化上高度保守。序列分析表明,ZmPP6C基因開放閱讀框?yàn)?12個(gè)核苷酸,編碼303個(gè)氨基酸殘基,包含PP2A的催化亞基PP2Ac結(jié)構(gòu)域;系統(tǒng)進(jìn)化樹分析表明,PP6C蛋白在進(jìn)化上較為保守,無論是單子葉植物還是雙子葉植物,氨基酸序列高度相似,可達(dá)91%。并且與高粱的PP6C蛋白相似性更高,高達(dá)99.67%。2.ZmPP6C基因在玉米葉片中表達(dá)量較高,并且響應(yīng)不同光質(zhì)處理。對玉米自交系B73的ZmPP6C基因進(jìn)行器官特異性表達(dá)分析表明,其表達(dá)量在成株期葉片中最高,是根中的7.9倍,在莖、雄花、葉枕、葉鞘和花柄中的表達(dá)量相對較低;ZmPP6C能夠響應(yīng)不同光質(zhì)處理,且受遠(yuǎn)紅光和紅光的影響較大。ZmPP6C的表達(dá)量在黑暗轉(zhuǎn)入遠(yuǎn)紅光30分鐘達(dá)到峰值,然后迅速下降,直到24 h一直保持較低的水平;轉(zhuǎn)入紅光15分鐘達(dá)到峰值,迅速下降后緩慢上升,直到24 h達(dá)到黑暗的7.2倍。3.ZmPP6C基因轉(zhuǎn)錄豐度能響應(yīng)長日照和短日照處理,暗示其可能通過光信號(hào)轉(zhuǎn)導(dǎo)通路來調(diào)控植物開花。在長日照條件下,玉米ZmPP6C的表達(dá)在長日條件下的光照和黑暗階段各有一個(gè)明顯的表達(dá)高峰,分別出現(xiàn)在進(jìn)入光照階段后10 h和進(jìn)入黑暗階段5 h時(shí)。在短日照條件下,ZmPP6C基因的表達(dá)豐度波動(dòng)起伏較大,最高峰值發(fā)生在進(jìn)入黑暗后10 h,次高峰值發(fā)生在進(jìn)入光照后8 h時(shí)。4.ZmPP6C基因轉(zhuǎn)錄豐度受脅迫處理的調(diào)節(jié)。在高滲透、鹽漬和淹水脅迫過程中,ZmPP6C基因的表達(dá)水平先降后升。B73幼苗在200 m M NaCl脅迫處理12 h、24 h和48 h時(shí),ZmPP6C的轉(zhuǎn)錄豐度是未處理的0.8倍、1.1倍和2.6倍。B73幼苗在20%PEG6000處理20 h時(shí),ZmPP6C基因的表達(dá)水平與未處理的相當(dāng);48 h時(shí),ZmPP6C的表達(dá)水平上升到未處理的的4.5倍。淹水處理處理5 d和7 d,ZmPP6C的表達(dá)分別上升到未處理的1.3倍和4.1倍;然后恢復(fù)正常溫室生長條件3 d和7 d,ZmPP6C的表達(dá)上升到未處理的3.0倍和6.9倍。表明ZmPP6C基因參與了玉米脅迫的應(yīng)答。
[Abstract]:Serine / threonine protein phosphatase 6 (catalytic subunits of Serr protein phosphatase 6 PP6C) is the catalytic subunit of PP6 whole enzyme. Studies in Arabidopsis thaliana showed that PP6C was involved in polar transport of auxin, abscisic acid signal transduction and light signal transduction mediated flowering regulation. In this study, the ZmPP6C gene was cloned by RT-PCR, and the evolutionary relationship between the protein structure and the homologous proteins was analyzed. The effects of different light quality, dark to various light quality, long sunlight and short sunlight were analyzed by quantitative PCR. And the expression pattern under stress, the result is as follows: 1. The amino acid sequence of protein phosphatase 6 subunit is highly conserved in evolution. Sequence analysis showed that the open reading frame of ZmPP6C gene was 912 nucleotides, encoding 303 amino acid residues, containing the catalytic subunit PP2Ac domain of PP2A, and phylogenetic tree analysis showed that PPP6C protein was more conserved in evolution. The amino acid sequences of monocotyledonous and dicotyledonous plants are similar, up to 91. The PP6C protein of sorghum was more similar to that of sorghum, as high as 99.67.2.ZmPP6C gene was expressed in maize leaves and responded to different light quality treatments. The organ-specific expression of ZmPP6C gene in maize inbred line B73 was analyzed. The results showed that the expression of ZmPP6C gene was the highest in adult leaves, 7.9 times higher than that in root, and 7.9 times in stem, male flower and leaf pillow. The expression of ZmPP6C in leaf sheath and petiole was relatively low, and the expression of ZmPP6C was significantly affected by far red light and red light. The expression of ZmPP6C reached its peak value at 30 minutes after the dark light was transferred to the far red light, and then decreased rapidly. The transcriptional abundance of ZmPP6C gene reached a peak level at 24 h, reached a peak value at 15 minutes of red light, decreased rapidly and increased slowly, until 24 h after transfer to dark 7.2-fold. 3. ZmPP6C gene transcription abundance could respond to long sunlight and short day exposure. It suggests that it may regulate plant flowering through the light signal transduction pathway. Under the condition of long sunlight, the expression of ZmPP6C in maize had an obvious peak in the light and dark stage, which appeared at 10 h after the light stage and 5 h after the dark stage, respectively. The expression abundance of ZmPP6C gene fluctuated greatly under the condition of short sunlight, the highest peak occurred at 10 hours after darkness, and the second peak occurred at 8 h after exposure to light. 4. The transcription abundance of ZmPP6C gene was regulated by stress treatment. At high penetration, The expression level of ZmPP6C gene decreased at first and then increased under 200 mm NaCl stress. The transcription abundance of ZmPP6C was 1.1 times higher than that of untreated ZmPP6C at 24 h and 48 h after 200mm NaCl stress, and 2.6-fold. B73 was ZmPP6C when treated with 20b PEG6000 for 20 h. The transcriptional abundance of ZmPP6C was 1.1 times and 2.6 times higher than that of untreated seedlings at 20 min PEG6000 for 20 h. The expression level of ZmPP6C was 4.5 times higher than that of untreated ZmPP6C at 48 h. The expression of ZmPP6C increased to 1.3 times and 4.1 times of untreated after 5 days and 7 days of flooding respectively, and then increased to 3.0 and 6.9 times of untreated after 3 days and 7 days of normal greenhouse growth conditions. The results showed that ZmPP6C gene was involved in maize stress response.
【學(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S513;Q943.2

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1 原換換;玉米ZmPP6C基因的克隆及其響應(yīng)光和脅迫處理的表達(dá)分析[D];河南農(nóng)業(yè)大學(xué);2016年

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本文編號(hào)：2095298