【摘要】：以三角褐指藻為材料,以巖藻黃素的合成途徑中主要的六個酶:八氫番茄紅素合成酶、八氫番茄紅素脫氫酶、ζ-胡蘿卜素脫氫酶、胡蘿卜素異構(gòu)酶、番茄紅素β-環(huán)化酶和玉米黃素環(huán)氧酶為研究對象,研究高光照脅迫對三角褐指藻巖藻黃素合成代謝途徑中關(guān)鍵基因表達量以及巖藻黃素含量的變化規(guī)律。以光照強度為500μE/(m2·s),分別照射處理三角褐指藻3h、6h、12h,采用HPLC對巖藻黃素含量進行定量分析。隨著光照時間的延長,巖藻黃素含量呈現(xiàn)先上升后下降的趨勢;高光照6h樣品中巖藻黃素含量為1.85mg·g-1DW(dry cell weight),比高光照0h對照組中巖藻黃素含量增加了2.16倍。qRT-PCR分析結(jié)果表明玉米黃素環(huán)氧酶基因zep1和八氫番茄紅素合成酶基因pys轉(zhuǎn)錄本在高光照脅迫6h時豐度最高,與對照組相比提高了近2倍,基因表達量的變化表現(xiàn)出與巖藻黃素含量變化相一致的趨勢。高光照處理3h后,胡蘿卜素異構(gòu)酶基因crtiso3、八氫番茄紅素脫氫酶基因pds、ζ-胡蘿卜素脫氫酶基因zds的表達水平相比于對照組明顯降低,表現(xiàn)出應(yīng)對高光照脅迫的快速響應(yīng);番茄紅素β-環(huán)化酶基因lcyb、胡蘿卜素異構(gòu)酶基因crtiso1的表達水平則在高光照處理12h后明顯提高;與zep1和pys相比,lcyb和crtiso1基因的表達對高光照脅迫處理的響應(yīng)時間要滯后至少6h。本研究揭示了高光照脅迫處理下三角褐指藻中巖藻黃素含量及其合成途徑中關(guān)鍵基因的表達模式,表明zep1和pys基因表達水平與巖藻黃素合成量之間存在明顯的線性關(guān)系。
[Abstract]:In this paper, six main enzymes in the biosynthesis pathway of phycoflavin, including lycopene synthase, lycopene dehydrogenase, 味 -carotene dehydrogenase, carotene isomerase, were used as materials. Lycopene 尾 -cyclase and zeaxanthin cyclooxygenase were used to study the effect of high light stress on the expression of key genes and the content of phycoflavin in the biosynthesis pathway of algin triangularis. The content of fucoflavin was quantitatively analyzed by HPLC in the light intensity of 500 渭 E / (m ~ 2 ~ 2 s),) irradiation for 3 h ~ 6 h ~ (12 h) of Phaeophyllum triangularis. With the prolongation of illumination time, the content of alginin increased first and then decreased. The content of alginin in the samples exposed to high light for 6 h was 2.16 times higher than that in the control group at 0 h. The results of qRT-PCR showed that zep1 and pys transcripts of zeaxanthin cyclooxygenase gene and octahydrolycopene synthase gene were increased by 2.16 times. The abundance was the highest at 6h of high light stress. Compared with the control group, the gene expression increased nearly 2 times, and the change of gene expression showed the same trend as the alginin content. The expression levels of carotene isomerase gene crtiso3, octahydrolycopene dehydrogenase gene pdsand 味 -carotene dehydrogenase gene zds were significantly lower than those of the control group after 3 h of high light exposure, showing a rapid response to high light stress. The expression of lycopene 尾 -cyclase gene lcyb and carotene isomerase gene crtiso1 increased significantly after 12 h of high light exposure, and the response time of lcyb and crtiso1 gene expression to high light stress was delayed by at least 6 hours compared with that of zep1 and pys. This study revealed the expression pattern of the key genes in the algin content and biosynthesis pathway of Phaeophyllum triangularis under high light stress, indicating that there was a significant linear relationship between the expression level of zep1 and pys genes and the amount of synthesizing of algin.
【作者單位】： 湖北大學生命科學學院;中國農(nóng)業(yè)科學院油料作物研究所農(nóng)業(yè)部油料作物生物學與遺傳重點實驗室;
【基金】：中國農(nóng)業(yè)科學院科技創(chuàng)新工程項目(CAAS-ASTIP-2013-OCRI)
【分類號】：Q946

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