【摘要】：獨行菜(Lepidium apetalum Wild.)和抱莖獨行菜(Lepidium perfoliatum L.)是十字花科獨行菜屬近緣種植物,為新疆具有代表性的早春短命植物。本課題組前期研究發(fā)現(xiàn)二者萌發(fā)及幼苗生長階段對低溫的耐受性有明顯差異,為進一步了解兩種獨行菜在種子休眠和萌發(fā)階段的差異性,本研究對二者種子低溫層積過程中生理生化特性進行了初步分析,并建立了適用于兩種獨行菜種子基因差異表達研究的序列相關擴增多態(tài)性分析體系(SRAP)。1比較了低溫層積處理對兩種獨行菜種子萌發(fā)特性的影響,對層積過程中過氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)、過氧化氫酶(catalase,CAT)活性的變化進行了測定。結果顯示:抱莖獨行菜種子萌發(fā)依賴低溫層積,而層積處理僅提高了獨行菜種子的萌發(fā)勢;兩種種子低溫層積過程中相關酶活性均發(fā)生變化,但變化趨勢不同。實驗結果提示兩種獨行菜種子對低溫層積處理具有不同的響應。此外,抱莖獨行菜種子不同部位提取物處理白菜種子的萌發(fā)實驗結果說明抱莖獨行菜種子休眠機制可能與其種胚中存在萌發(fā)抑制物有關。2利用高效液相色譜測定了兩種獨行菜種子低溫層積過程中赤霉素(GA)和脫落酸(ABA)含量的變化情況。結果顯示,二者在層積過程中種子內源激素變化趨勢相同:GA含量上升,ABA含量下降;抱莖獨行菜種子層積處理前ABA濃度較高,獨行菜種子層積處理前GA濃度較高,且獨行菜種子在層積過程中激素變化幅度較大。結果提示GA和ABA在低溫層積處理解除兩種獨行菜種子的休眠過程中可能具有重要作用。3研究了外源激素處理對兩種獨行菜種子休眠與萌發(fā)的影響。IAA、NAA可部分解除抱莖獨行菜的種子休眠,對獨行菜種子萌發(fā)則表現(xiàn)抑制作用。GA對兩種種子均促進萌發(fā),并且可完全解除抱莖獨行菜種子的休眠。6-BA對解除抱莖獨行菜種子休眠無作用,對獨行菜種子萌發(fā)表現(xiàn)兩重性。ABA嚴重抑制兩種種子萌發(fā)。結果顯示兩種獨性菜種子休眠釋放過程對激素的響應各有異同。4為建立適合兩種獨行菜種子的cDNA-SRAP反應體系,分析了dNTPs、Mg2+、引物、cDNA模板、Taq DNA聚合酶的濃度對擴增結果的影響,并對擴增條件進行了優(yōu)化。最終確定優(yōu)化的cDNA-SRAP反應體系為:20μL體系中cDNA模板100 ng,Mg2+濃度1.75 mmol·L-1,dNTPs濃度0.275 mmol·L-1,引物濃度1.0μmol·L-1,Taq DNA聚合酶0.5 U。該體系擴增條帶數(shù)目較多且較清晰,可用于后續(xù)低溫層積處理下兩種獨行菜基因差異表達的研究。
[Abstract]:Lepidium apetalum Wild. And Lepidium perfoliatum L. It is a relative species of Cruciferae, which is a typical short-lived early spring plant in Xinjiang. The preliminary study of our group found that there were significant differences in the tolerance to low temperature between the two species during germination and seedling growth, in order to further understand the differences between the two species in seed dormancy and germination stage. In this study, the physiological and biochemical characteristics of the seeds during their low temperature stratification were preliminarily analyzed. A sequence-dependent amplified polymorphism analysis system (SRAP) was established for gene differential expression analysis of two species of single vegetable seeds. 1. The effects of low temperature stratification on seed germination characteristics were compared. The activities of peroxidase (POD), superoxide dismutase (superoxide) and catalase (cat) during stratification were determined. The results showed that the seed germination was dependent on low temperature stratification, but the seed germination potential was only increased by stratification, and the related enzyme activities of the two seeds changed during the low temperature stratification, but the change trend was different. The results showed that the two seeds had different responses to low temperature stratification. In addition, Germination of Chinese Cabbage Seeds treated with different extracts from Stem Seeds. 2. The dormancy mechanism of Chinese cabbage seeds may be related to the existence of germination inhibitors in their embryos. 2. Two kinds of seeds were determined by high performance liquid chromatography (HPLC). Changes of gibberellin (GA) and abscisic acid (ABA) contents during the low temperature stratification of the seeds. The results showed that the change trend of endogenous hormones in seeds was the same during stratification, the content of GA increased and the content of ABA decreased, while the concentration of ABA was higher before stratification and higher in GA before stratification. In addition, the changes of hormone in the seed of monopodium chinensis were larger during the stratification process. The results suggest that GA and ABA may play an important role in the dormancy process of the seeds of two species of single vegetable treated by low temperature stratification. 3. The effects of exogenous hormone treatment on the dormancy and germination of the seeds of the two species were studied. The seed dormancy of the single stem, On the other hand, GA promoted the germination of both seeds, and completely relieved the dormancy of the seeds. 6-BA had no effect on the dormancy of the seeds. The germination of the two seeds showed duality. ABA seriously inhibited the germination of the two kinds of seeds. The results showed that there were similarities and differences in the responses to hormones in the dormancy release process of the two species. 4. In order to establish a cDNA-SRAP reaction system suitable for the two kinds of seeds, the effects of the concentration of dNTPsMg2, primer cDNA template Taq DNA polymerase on the amplification results were analyzed. The amplification conditions were optimized. The optimized cDNA-SRAP reaction system was determined as follows: the concentration of 100ng / mg _ 2 and primer was 1.75 mmol / L ~ (-1), 0.275 mmol / L ~ (-1) and 1.0 渭 mol / L ~ (Taq) Taq DNA polymerase in 1: 20 渭 L / L system respectively. This system can be used to study the gene differential expression of two kinds of monopodium under the following low temperature stratification treatment because of the large number of amplified bands and the clarity of the amplified bands.
【學位授予單位】：新疆大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q945.3;Q943.2

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