本文選題：辣椒 + 果實(shí)　； 參考：《中國農(nóng)業(yè)大學(xué)》2015年博士論文

【摘要】：從一個(gè)辣椒RIL (recombinant inbred line,重組自交系)群體中選出果實(shí)大小和形狀各異的49份材料,連同雙親及雜交一代共52份材料為試材,對子房和成熟果實(shí)進(jìn)行細(xì)胞學(xué)研究發(fā)現(xiàn),在辣椒果皮橫切和縱切面上,果皮(子房壁)的結(jié)構(gòu)很保守,在授粉之前已經(jīng)確定,果皮細(xì)胞可以清晰的劃分為：外果皮、中果皮和內(nèi)果皮細(xì)胞。相關(guān)性分析表明,果實(shí)橫徑的生長模式比較保守；除細(xì)胞形狀外,細(xì)胞數(shù)目也在很大程度上影響果實(shí)形狀。從果實(shí)的宏觀生長來看,果實(shí)橫徑與果皮厚度正相關(guān),果實(shí)縱徑與二者不相關(guān),呈現(xiàn)出異速增長模式,形成了果實(shí)形狀的多樣性。果皮細(xì)胞的生長模式與果實(shí)宏觀生長相似,也表現(xiàn)出異速增長模式。 以羊角形辣椒自交系“0819”為試材,研究了果實(shí)生長過程中外部形態(tài)和細(xì)胞的動態(tài)變化。“0819”果實(shí)生長過程中,果實(shí)縱徑、橫徑和果皮厚度具有相似的“S”形生長曲線,在縱向和橫向切面上,中果皮細(xì)胞平均面積增長曲線也大致呈“S”形。細(xì)胞徑向分裂主要發(fā)生在果實(shí)生長早期的最外層細(xì)胞,以增大果實(shí)橫切面周長適應(yīng)中果皮細(xì)胞的膨大；在10DAP (days after pollination,授粉后天數(shù))之前,外果皮及中果皮細(xì)胞進(jìn)行活躍的橫向分裂,是果實(shí)縱徑伸長的重要影響因素；平周分裂主要發(fā)生在外層亞表皮細(xì)胞層,大約在20DAP停止。細(xì)胞膨大主要發(fā)生在中果皮,并對果實(shí)橫徑、果皮厚度以及后期的果實(shí)縱徑伸長起決定作用。 采用ELISA方法,對0、5、10、50DAP果實(shí)中IAA、 GA、 ZR和ABA的含量進(jìn)行測定,發(fā)現(xiàn)種子和胎座中激素含量的變化趨勢大致相同,而果皮中的變化趨勢與之有明顯差異。在5DAP時(shí)辣椒種子中赤霉素含量升高,生長素含量降低,而在5DAA-NP時(shí)種子生長素含量仍然較高,赤霉素含量降低,說明赤霉素可能是促進(jìn)辣椒果實(shí)坐果的主要激素,而且授粉受精后種子中赤霉素的合成并非與生長素的大量合成相關(guān)。 對0、5、10、50DAP果實(shí)進(jìn)行轉(zhuǎn)錄組測序,發(fā)現(xiàn)在5和10DAP專一表達(dá)的基因數(shù)目比0DAP和50DAP專一表達(dá)的基因數(shù)目少,說明5和10DAP果實(shí)主要進(jìn)行膨大生長,生長狀態(tài)相似,而0DAP和50DAP時(shí),果實(shí)(子房)進(jìn)行著更為復(fù)雜的反應(yīng)；植物激素相關(guān)基因的表達(dá)也顯示,5DAP和10DAP的果實(shí)基因的表達(dá)模式相似。 同源克隆了3個(gè)赤霉素受體基因CaGIDlb.1、 CaGIDlb.2和CaGID1c,以及一個(gè)赤霉素信號轉(zhuǎn)導(dǎo)途徑中的負(fù)調(diào)控因子DELLA蛋白基因CaGAI。通過基因表達(dá)定量分析,發(fā)現(xiàn)在辣椒坐果過程中,CaGIDlb.2在果皮和胎座中發(fā)揮著重要作用；而CaGIDlb.／則在種子中可能有更重要的作用。在果實(shí)生長發(fā)育的過程中,CaGIDIb.l和CaGIDlb.2可能比CaGIDlc有更重要的作用。將CaGIDlb.1、 CaGIDlb.2和CaGID1c在擬南芥雙突變體gidlagidlc中過表達(dá),株高顯著增加；將CaGAI在擬南芥雙突變體rga-24/gai-t6中過表達(dá),株高降低,表明CaGIDls和CaGA1影響植株莖的伸長,與擬南芥中AtGID1和AtGAI的作用相似。酵母雙雜(Y2H)和熒光雙分子互補(bǔ)(BiFC)試驗(yàn)表明,在GA3存在的條件下,CaGID1b1和CaGID1b2可以與CaGAI相互作用；而CaGID1c與CaGAI的相互作用不依賴于GA3。
[Abstract]:49 samples of different sizes and shapes of fruit were selected from a group of RIL (recombinant inbred line, recombinant inbred line). Together with 52 copies of a total of two parents and hybrids, cytological studies on the ovary and mature fruit found that the structure of the peel (ovary wall) was conservative in the cross and longitudinal section of Capsicum. Before the powder was determined, the peel cells could be clearly divided into the outer skin, the pericarp and the endocarp cells. The correlation analysis showed that the growth pattern of the transverse diameter of the fruit was more conservative; the cell number also influenced the shape of the fruit to a great extent except the shape of the cells. The longitudinal diameter of fruit is not related to the two, showing a pattern of different speed growth, which forms the diversity of fruit shape. The growth pattern of fruit skin cells is similar to the macro growth of fruit, and also shows a pattern of different speed growth.
The dynamic changes of external morphology and cells in the growth process of the capsicum pepper inbred line 0819 were studied. "0819" fruit growth process, the fruit longitudinal diameter, transverse diameter and pericarp thickness have similar "S" shape growth curve, and the average area growth curve of the pericarp cells in the longitudinal and transverse sections is also roughly presented. "S" shape. Cell radial division mainly occurs in the outermost cells of the early growth of the fruit, which increases the expansion of the pericarp cells in the transverse section of the fruit. Before the 10DAP (days after pollination, the number of days after pollination), the active transverse division of the outer skin and the medium pericarp cells is an important factor affecting the lengthwise elongation of the fruit. The division of the circumferential Division mainly occurs in the outer layer of the subepidermal cell layer and stops at about 20DAP. The cell expansion mainly occurs in the pericarp, which determines the transverse diameter of the fruit, the thickness of the pericarp and the lengthwise elongation of the later fruit.
ELISA method was used to determine the content of IAA, GA, ZR and ABA in the fruits of 0,5,10,50DAP. It was found that the change trend of hormone content in seed and placenta was approximately the same, but there was a significant difference in the pericarp. At 5DAP, the content of gibberellin and the content of auxin in the pepper seeds were lower, while the seed auxin was contained in 5DAA-NP. The amount of gibberellin may be the main hormone to promote fruit sitting of capsicum, and the synthesis of gibberellin in seed after pollination and fertilization is not related to the synthesis of auxin.
The transcriptional sequence of 0,5,10,50DAP fruit showed that the number of genes expressed in 5 and 10DAP was less than that of 0DAP and 50DAP, indicating that 5 and 10DAP fruits were mainly expanded, and the growth state was similar, while the fruit (Zi Fang) was more complex when 0DAP and 50DAP, and the expression of plant hormone related genes was expressed. It also showed that the expression patterns of 5DAP and 10DAP were similar.
Homologous cloning of 3 gibberellin receptor genes CaGIDlb.1, CaGIDlb.2 and CaGID1c, and a negative regulator of gibberellin signal transduction pathway DELLA protein gene CaGAI. through quantitative analysis of gene expression, it is found that CaGIDlb.2 plays an important role in the peel and placenta during the fruit setting of capsicum, and CaGIDlb. / in the seed. In the process of fruit growth, CaGIDIb.l and CaGIDlb.2 may have a more important role than CaGIDlc. CaGIDlb.1, CaGIDlb.2 and CaGID1c are overexpressed in the Arabidopsis double mutant gidlagidlc, and the height of the plant increases significantly; CaGAI is overexpressed in the Arabidopsis thaliana double mutant rga-24/gai-t6, and the height of the strain is reduced. The effect of CaGIDls and CaGA1 on the elongation of plant stems is similar to that of AtGID1 and AtGAI in Arabidopsis. Yeast double heterozygosity (Y2H) and fluorescent double molecular complementation (BiFC) test show that CaGID1b1 and CaGID1b2 can interact with CaGAI under the presence of GA3, while the interaction between CaGID1c and CaGAI is not dependent on the CaGAI.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S641.3

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