發(fā)布時間：2018-05-18 16:00

  本文選題：柑橘 + 枳��； 參考：《華中農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：硼是高等植物生長發(fā)育所必需的微量元素。我國大多數(shù)柑橘種植區(qū)的土壤含硼量低,柑橘缺硼現(xiàn)象比較普遍。柑橘一般采用嫁接繁殖,砧木根系是柑橘對養(yǎng)分吸收和轉(zhuǎn)運的基礎(chǔ),研究缺硼脅迫對砧木根系生長發(fā)育的影響對于解決柑橘生產(chǎn)中的缺硼問題具有重要意義。本研究選取柑橘生產(chǎn)上常用的砧木—枳[Poncirus trifoliate(L.)Raf.]為研究材料,采用Hogland營養(yǎng)液水培的方法,從形態(tài)、生理、解剖和分子幾方面來研究缺硼對枳根系生長發(fā)育的影響。主要結(jié)果如下:1.缺硼對枳根系形態(tài)和解剖結(jié)構(gòu)的影響缺硼處理90d,植株的主根和側(cè)根的伸長明顯受到抑制,根系表現(xiàn)為粗短叢枝狀,根尖膨大甚至壞死,整個根系顏色加深偏老齡化,根尖伸長區(qū)和分生區(qū)的長度明顯縮短,側(cè)根往根尖靠近。缺硼植株的側(cè)根密度和側(cè)根原基的密度都顯著大于對照。石蠟切片觀察發(fā)現(xiàn):缺硼導(dǎo)致枳根尖明顯膨大變粗,細胞數(shù)量增多,細胞排列混亂無序,無法觀察到明顯的靜止中心,根尖生長點壞死,嚴(yán)重時壞死細胞脫落形成空腔,有大量的維管束增生,根表皮破裂。枳根尖縱切掃描電鏡觀察發(fā)現(xiàn),缺硼植株根尖細胞壁有增厚,細胞間隙增大,細胞排列不緊湊。透射電鏡觀察發(fā)現(xiàn)缺硼根尖細胞核靠壁固縮變形,液泡極度增大,細胞器消失,細胞壁之間的膠連瓦解,細胞壁明顯加厚,細胞壁內(nèi)壁有物質(zhì)積累,導(dǎo)管內(nèi)壁也有大量的物質(zhì)積累。2.缺硼對枳根中細胞周期相關(guān)基因以及根系伸長相關(guān)基因表達的影響根系的生長需要兩個條件:一是根尖分生區(qū)細胞不斷分裂增生;二是根伸長區(qū)細胞伸長生長。因此,我們篩選了幾個與細胞周期調(diào)控相關(guān)的基因PtcycA,PtcycB,PtcycD3和Ptcdc2和根系伸長相關(guān)的基因PtLRP1和PtSWP1來做表達分析,觀察缺硼脅迫對它們表達模式的影響。其結(jié)果顯示:細胞周期相關(guān)基因的表達模式受缺硼脅迫的影響不大。PtLRP1基因在缺硼處理18h前表達量明顯上調(diào),但缺硼處理48h后,其表達量急劇下降,且明顯低于對照。PtSWP1基因表達量從缺硼處理18h開始上調(diào)表達,而且其表達量顯著高于對照,最高的時候是對照的3倍以上。3.缺硼對枳根中IAA含量的影響對缺硼處理90d后的枳植株添加0.3mg/L的外源IAA,結(jié)果發(fā)現(xiàn):在有外源IAA存在的情況下,缺硼植株又能重新長出很多新根。新根根尖的解剖結(jié)構(gòu)與正常硼營養(yǎng)下的植株根尖相比,除了沒有正常根尖長得勻稱和成熟區(qū)更接近根尖外,其它都比較正常,都能觀察到比較明顯的靜止中心,有明顯的根冠。短期缺硼處理對枳根中束縛態(tài)硼的濃度影響不大,可溶性硼的濃度隨著缺硼處理的時間延長而逐漸降低。在缺硼條件下,枳根中IAA的含量與可溶性硼的濃度有相同的下降趨勢。IAA的含量在缺硼處理24h前顯著升高,且明顯高于對照;隨著處理時間延長,到第48h后,枳根中IAA含量急劇下降,且顯著低于對照根中的IAA含量。4.缺硼對枳根中IAA代謝相關(guān)基因表達的影響為了進一步明確缺硼是怎樣影響枳根中IAA的含量,選擇了一些IAA代謝相關(guān)的基因來做表達分析,觀察缺硼對這些基因表達的影響。實驗結(jié)果發(fā)現(xiàn):生長素合成相關(guān)基因TAA1,TAR2,YUC3和YUC8在缺硼處理12h前明顯被誘導(dǎo)表達,12h后這些基因的表達量顯著降低,直到實驗結(jié)束。這些基因的表達模式與根中IAA含量的變化一致。另外,在缺硼情況下,向基性轉(zhuǎn)運蛋白基因AUX1,PIN1和PIN4的表達水平明顯下調(diào),而向頂性轉(zhuǎn)運蛋白基因LAX1,ABCB1和PIN3的表達水平顯著升高,說明流向根尖的IAA量減少,而從根尖向外運出的IAA量增加。
[Abstract]:Boron is a necessary trace element for the growth and development of higher plants. The boron content of the soil in most of the citrus planting areas in China is low, and the phenomenon of boron deficiency in citrus is common. The root of the citrus is the basis for the absorption and transport of the citrus. The effect of boron deficiency on the growth and development of the root of the anvil system is studied to solve the citrus students. The problem of boron deficiency in the production is of great significance. In this study, we selected the stock of orange [Poncirus trifoliate (L.) Raf.] as the research material, and used the method of Hogland nutrient solution hydroponics to study the effects of boron deficiency on the growth and development of the roots of trifoliate orange from the morphological, physiological, anatomical and molecular aspects. The main results are as follows: 1. the deficiency of boron to the trifoliate orange is the main result. The effect of root morphology and anatomical structure on 90d, the elongation of the main root and the lateral root of the plant was obviously inhibited, the root of the root showed a coarse and short branching, the root tip expanded or even necrotic. The root color deepened, the length of the root tip elongated and the sub zone shortened obviously, the lateral root was close to the root tip. The lateral root density of the boron deficient plant was the root density and the root density of the boron deficient plant. The density of the lateral root primordium was significantly greater than that of the control. The observation of paraffin section showed that the lack of boron resulted in the obvious enlargement and thickening of the root tip of the trifoliate, the number of cells increased, the cells arranged disorder and disorder, the obvious static center could not be observed, the tip of the root tip was necrotic, and the necrotic cells fall off into the cavity, and a large number of vascular bundles were proliferated and the root epidermis ruptured. It was found that the cell wall of the root tip of the boron deficient plant was thickened, the cell gap increased and the cell arrangement was not compact. The transmission electron microscope observed that the nucleus of the root apex of boron deficiency was deformed, the vacuole was greatly increased, the organelle disappeared, the cell wall was thickened and the cell wall was thickened, and the inner wall of the cell wall was accumulated. There are also a large amount of material accumulation in the inner wall of the catheter, which affects the growth of the cell cycle related genes and the expression of root elongation related genes in the root of the trifoliate orange. Two conditions are required for the growth of the root system: one is the cell proliferation in the apical sub region, and the two is the growth of the cells in the root elongation region. Therefore, we screened several regulatory phases with the cell cycle. The gene PtcycA, PtcycB, PtcycD3 and Ptcdc2 and the genes related to root elongation, PtLRP1 and PtSWP1, were used for expression analysis to observe the effect of boron deficiency on their expression patterns. The results showed that the expression pattern of cell cycle related genes was not affected by boron deficiency stress, but the expression of.PtLRP1 gene was up significantly up before the boron deficiency treated 18h. After boron deficiency, the expression of 48h decreased dramatically, and the expression of.PtSWP1 gene expression was significantly lower than that of the control 18h, and the expression amount was significantly higher than that of the control. At the highest level, the.3. deficiency was more than 3 times that of the control. The effect of.3. deficiency on the IAA content in the root of trifoliate orange was added to the exogenous IAA in the 0.3mg/L after 90d deficiency. It was found that in the presence of exogenous IAA, the boron deficient plant could grow a lot of new roots again. The anatomical structure of the root tip of the new root is more normal than the normal root tip and the mature area is closer to the root tip than that of the plant root tip under the normal boron nutrition. The short-term boron deficiency treatment had little effect on the concentration of bound boron in the root of trifoliate orange, and the concentration of soluble boron gradually decreased with the prolongation of the time of boron deficiency. In the condition of boron deficiency, the content of IAA and the concentration of soluble boron in the root of the citrus trifoliate had the same decreasing trend as the content of.IAA, which was significantly higher before the boron deficiency treatment 24h, and obviously higher than that of the boron treatment. With the prolongation of treatment time and 48h, the content of IAA in the root of trifoliate orange decreased sharply and was significantly lower than the IAA content in the control root. The effect of boron deficiency on the expression of IAA metabolism related genes in the root of trifoliate trifoliate (trifoliate orange) in order to further clarify the effect of boron deficiency on the content of IAA in the root of trifoliate trifoliate, selected some genes related to IAA metabolism and observed the expression analysis. The effect of boron deficiency on the expression of these genes. The experimental results showed that the gene TAA1, TAR2, YUC3 and YUC8 were obviously induced before 12h, and the expression of these genes decreased significantly until the end of the experiment. The expression patterns of these genes were consistent with the changes in the content of IAA in the root. In addition, in the case of boron deficiency, the gene expression pattern was in the absence of boron. The expression level of the basic transporter gene AUX1, PIN1 and PIN4 was obviously down, while the expression level of LAX1, ABCB1 and PIN3 increased significantly, indicating the decrease of IAA in the root tip and the increase of IAA from the root tip.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S666

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