辣椒果實細(xì)胞學(xué)、轉(zhuǎn)錄組分析及赤霉素受體CaGID1s和DELLA蛋白CaGAI功能、互作研究
本文選題:辣椒 + 果實; 參考:《中國農(nóng)業(yè)大學(xué)》2015年博士論文
【摘要】:從一個辣椒RIL (recombinant inbred line,重組自交系)群體中選出果實大小和形狀各異的49份材料,連同雙親及雜交一代共52份材料為試材,對子房和成熟果實進行細(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形狀。從果實的宏觀生長來看,果實橫徑與果皮厚度正相關(guān),果實縱徑與二者不相關(guān),呈現(xiàn)出異速增長模式,形成了果實形狀的多樣性。果皮細(xì)胞的生長模式與果實宏觀生長相似,也表現(xiàn)出異速增長模式。 以羊角形辣椒自交系“0819”為試材,研究了果實生長過程中外部形態(tài)和細(xì)胞的動態(tài)變化!0819”果實生長過程中,果實縱徑、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)生在果實生長早期的最外層細(xì)胞,以增大果實橫切面周長適應(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)胞進行活躍的橫向分裂,是果實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,并對果實橫徑、果皮厚度以及后期的果實縱徑伸長起決定作用。 采用ELISA方法,對0、5、10、50DAP果實中IAA、 GA、 ZR和ABA的含量進行測定,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時辣椒種子中赤霉素含量升高,生長素含量降低,而在5DAA-NP時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辣椒果實坐果的主要激素,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果實進行轉(zhuǎn)錄組測序,發(fā)現(xiàn)在5和10DAP專一表達的基因數(shù)目比0DAP和50DAP專一表達的基因數(shù)目少,說明5和10DAP果實主要進行膨大生長,生長狀態(tài)相似,而0DAP和50DAP時,果實(子房)進行著更為復(fù)雜的反應(yīng);植物激素相關(guān)基因的表達也顯示,5DAP和10DAP的果實基因的表達模式相似。 同源克隆了3個赤霉素受體基因CaGIDlb.1、 CaGIDlb.2和CaGID1c,以及一個赤霉素信號轉(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,株高顯著增加;將CaGAI在擬南芥雙突變體rga-24/gai-t6中過表達,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(BiFC)試驗表明,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
【參考文獻】
相關(guān)期刊論文 前9條
1 符特;羅新蘭;李天來;任樂;花妍;;日光溫室番茄果實鮮重與橫縱徑數(shù)學(xué)關(guān)系研究[J];北方園藝;2007年05期
2 鄭家基;;橄欖生長與結(jié)果習(xí)性的觀察研究[J];福建農(nóng)學(xué)院學(xué)報;1988年02期
3 陳清西,廖鏡思,胡又厘;龍眼果實生長曲線和各組織的相關(guān)分析[J];福建農(nóng)業(yè)大學(xué)學(xué)報;1995年01期
4 王剛;蘭竹荔枝生長及果實發(fā)育規(guī)律研究[J];貴州農(nóng)業(yè)科學(xué);1994年06期
5 閻立英;草莓果實生長曲線的研究[J];河北林果研究;1998年S1期
6 晏儒來,姚明華;茄子果實及種子生長發(fā)育的特點[J];湖北農(nóng)業(yè)科學(xué);1993年10期
7 王立浩;毛勝利;張正海;張寶璽;;辣椒分子遺傳育種研究進展[J];辣椒雜志;2012年01期
8 周華;張新;劉騰云;余發(fā)新;;高通量轉(zhuǎn)錄組測序的數(shù)據(jù)分析與基因發(fā)掘[J];江西科學(xué);2012年05期
9 李明,郝建軍,于洋,付淑杰,李云鵬,吳建麗;脫落酸(ABA)對蘋果果實著色相關(guān)物質(zhì)變化的影響[J];沈陽農(nóng)業(yè)大學(xué)學(xué)報;2005年02期
,本文編號:2038377
本文鏈接:http://sikaile.net/shoufeilunwen/nykjbs/2038377.html