本文選題：條斑紫菜 + 非生物脅迫��； 參考：《中國科學(xué)院研究生院(海洋研究所)》2016年博士論文

【摘要】：條斑紫菜是一種重要的經(jīng)濟海藻。近年來條斑紫菜種質(zhì)退化,影響了紫菜產(chǎn)業(yè)的發(fā)展,培育優(yōu)良的種質(zhì)成為亟需解決的問題。鑒于條斑紫菜生活環(huán)境的特點,可以從提高其抗性方面進行良種培育。良種培育需要有優(yōu)良性狀的種質(zhì)資源,但是條斑紫菜育種研究相對陸地作物來說起步較晚,基礎(chǔ)研究也比較薄弱,目前還沒有明確的與抗性特征相關(guān)的位點和分子標(biāo)記。正因如此,本論文聚焦條斑紫菜的抗性育種,一方面研究其光合作用過程對鹽脅迫條件的響應(yīng),確定藻體抗性特征的關(guān)鍵位點;另一方面基于條斑紫菜基因組測序信息開發(fā)SSR標(biāo)記,通過人工誘變擴展其遺傳多樣性,為定位抗性特征相關(guān)數(shù)量性狀位點奠定基礎(chǔ)。主要研究結(jié)果如下:(1)分析條斑紫菜光合作用過程對鹽脅迫的響應(yīng),發(fā)現(xiàn)光系統(tǒng)II(PSII)對高鹽脅迫條件比光系統(tǒng)I(PSI)更敏感。在高鹽(120‰)條件下,PSII的實際光合量子產(chǎn)量(effective PSII quantum yield,YII)幾乎為零,但是PSI的實際光合量子產(chǎn)量(effective PSI quantum yield,YI)維持較高活性,表明在PSII被抑制時PSI運轉(zhuǎn)時的電子來源于基質(zhì)側(cè)還原力。條斑紫菜隨著鹽度的升高藻體淀粉含量呈下降的趨勢,可溶性糖含量卻與其相反;同時,隨著鹽度的升高磷酸戊糖途徑(oxidative pentose phosphate pathway,OPPP)的關(guān)鍵酶6-磷酸葡萄糖脫氫酶(glucose-6-phosphate dehydrogenase,EC 1.1.1.49,G6PDH)和6-磷酸葡糖酸脫氫酶(6-phosphogluconate dehydrogenase,EC 1.1.1.44,6PGDH)的活性呈上升趨勢,糖酵解途徑中的重要酶3-磷酸甘油醛脫氫酶(cytosolic glyceraldehyde 3-phosphate dehydrogenase,EC 1.2.1.12,GAPDH)活性卻呈下降的趨勢。紫菜細胞還原力含量測定結(jié)果顯示,由OPPP途徑產(chǎn)生的NADPH含量呈升高的趨勢,而糖酵解途徑中產(chǎn)生的NADH含量卻呈下降趨勢。研究結(jié)果說明,在鹽脅迫的條件下淀粉降解速度加快,產(chǎn)生的可溶性糖進入OPPP途徑,產(chǎn)生的還原力NADPH推動了PSI的環(huán)式電子傳遞運行。(2)基于條斑紫菜全基因組測序信息,開發(fā)了700對完全重復(fù)類型的SSR標(biāo)記。在270 M基因組中存在254495個SSR標(biāo)記,平均每隔1 kb存在1個SSR標(biāo)記;條斑紫菜的SSR標(biāo)記包括2-6個堿基的重復(fù),其中兩堿基所占的比例最多,其次是三堿基。重復(fù)類型中CG/GC比例最高,其次是CGG/CCG類型。在保存的16個條斑紫菜樣品中分析了SSR標(biāo)記的多態(tài)性情況,發(fā)現(xiàn)98對SSR標(biāo)記在這些樣品中具有多態(tài)性。根據(jù)PCR產(chǎn)物電泳結(jié)果進行遺傳分析,發(fā)現(xiàn)平均等位基因數(shù)(Na)為2.479;有效等位基因數(shù)(Ne)為1.558;香農(nóng)信息指數(shù)(I)為0.543;觀察雜合度(Ho)為0.168,期望雜合度(He)為0.32,PIC為0.35。聚類分析將這16個樣品聚成了兩大類,說明種質(zhì)混雜,樣品間存在基因交流。(3)化學(xué)(EMS)和物理(重離子束輻照)方法對條斑紫菜絲狀體進行了誘變,結(jié)果表明:0.2 M EMS處理2 h、0.4 M EMS處理1.5 h和0.6 M EMS處理1h是比較有效的化學(xué)誘變方法;重離子束輻照方法表明100-150戈瑞(GY)是比較有效的劑量。隨機選取20個重離子束輻照絲狀體樣品,用SSR分子標(biāo)記分析其遺傳多樣性,結(jié)果表明:平均等位基因數(shù)為(Na)2.842;有效等位基因數(shù)(Ne)為1.935;香農(nóng)信息指數(shù)(I)為0.756;觀察雜合度(Ho)為0.656,期望雜合度(He)為0.46,PIC平均為0.39。誘變條斑紫菜聚類分析結(jié)果表明不同的樣品基本按照重離子束輻照時所用的劑量進行聚類,這些樣品被分成了幾個亞類,最終這些亞類同對照聚到一起。這些誘變的樣品較保存的條斑紫菜遺傳多樣性高。
[Abstract]:Porphyra yezoensis is an important economic seaweed. In recent years, the germplasm degradation of Porphyra yezoensis has affected the development of the Porphyra yezoensis industry. It is an urgent problem to cultivate fine germplasm. In view of the characteristics of the living environment of Porphyra yezoensis, it can be cultivated from the improvement of its resistance. The breeding research of Porphyra yezoensis is relatively late and the basic research is relatively weak. There are no specific loci and molecular markers related to resistance characteristics. This paper focuses on the resistance breeding of Porphyra yezoensis. On the one hand, it studies the response of the light cooperation process to the salt stress conditions and determines the resistance of the algae. On the other hand, based on the genome sequencing information of Porphyra yezoensis, SSR markers were developed to expand the genetic diversity by artificial mutagenesis to lay the foundation for the quantitative trait loci of the resistance characteristics. The main results are as follows: (1) the response of light cooperative process to salt stress of Porphyra yezoensis was analyzed, and the light system II (PS) was found. II) the high salt stress condition is more sensitive than the light system I (PSI). Under the condition of high salt (120 per thousand), the actual photosynthetic quantum yield of PSII (effective PSII quantum yield, YII) is almost zero, but the actual photosynthetic quantum yield of PSI (effective PSI) is highly active. With the increase of salinity, the starch content of the algae declined with the increase of salinity, but the content of soluble sugar was opposite. At the same time, with the increase of salinity, the key enzyme of pentose phosphate pathway (oxidative pentose phosphate pathway, OPPP) was 6- phosphoric glucose dehydrogenase (glucose-6-phosphate dehydrogenase, EC 1.1.1.49, G6P) DH) and 6- phosphate dehydrogenase (6-phosphogluconate dehydrogenase, EC 1.1.1.44,6PGDH) are on the rise. The activity of 3- phosphate glyceraldehyde dehydrogenase (cytosolic glyceraldehyde 3-phosphate dehydrogenase, EC) is declining. The results showed that the content of NADPH produced by OPPP pathway increased and the content of NADH produced in the glycolysis pathway decreased. The results showed that the degradation rate of starch was accelerated under salt stress, the soluble sugar produced into the OPPP pathway, and the resulting reducing force NADPH promoted the ring type electron transfer operation of PSI. (2) based on the results, the results showed that the degradation rate of soluble sugar produced by the glycolysis was accelerated. The whole genome sequencing information of Porphyra yezoensis has developed 700 pairs of fully duplicated SSR markers. There are 254495 SSR markers in the 270 M genome, with an average of 1 SSR markers per 1 KB; the SSR marker of Porphyra yezoensis includes 2-6 base repeats, among which two bases occupy the largest proportion, followed by the three base. The CG/GC ratio in the repeat type. The highest, followed by the CGG/CCG type. The polymorphism of the SSR markers was analyzed in the 16 preserved Porphyra samples, and 98 pairs of SSR markers were found to be polymorphic in these samples. The average number of alleles (Na) was 2.479, the number of effective alleles (Ne) was 1.558, and the Shannon information index (I) was found. ) for 0.543, the observation heterozygosity (Ho) was 0.168, the expected heterozygosity (He) was 0.32, and PIC was 0.35. clustering analysis to gather the 16 samples into two categories, indicating that the germplasm was mixed and the gene exchange existed between the samples. (3) chemical (EMS) and Physics (heavy ion beam irradiation) were used to mutagenate the filamentous body of Porphyra yezoensis. The results showed that 0.2 M EMS treatment 2 h, 0.4 M E. MS processing 1.5 h and 0.6 M EMS to treat 1H is a more effective chemical mutagenesis method; heavy ion beam irradiation method shows that 100-150 Gore (GY) is a more effective dose. 20 heavy ion beams are randomly selected to irradiate filamentous samples and the genetic diversity is analyzed with SSR molecular markers. The results show that the average allele number is (Na) 2.842, and the effective allele is effective. The factor (Ne) was 1.935, the Shannon information index (I) was 0.756, the observation heterozygosity (Ho) was 0.656, the expected heterozygosity (He) was 0.46, and the PIC was 0.39. mutagenesis. The cluster analysis of Porphyra yezoensis showed that the different samples were basically clustered according to the dose used in the heavy ion beam irradiation, and these samples were divided into several subclasses, and finally these subcategories were similar. Compared with the preserved Porphyra yezoensis, these mutagenic samples had higher genetic diversity.
【學(xué)位授予單位】：中國科學(xué)院研究生院(海洋研究所)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S917.3

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