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      桃果皮葉綠素降解規(guī)律與相關(guān)基因表達(dá)分析

      2024-06-30 02:07:38劉鑫張曉煜孟君仁段文宜孫世航潘磊曾文芳王志強(qiáng)牛良
      果樹(shù)學(xué)報(bào) 2024年6期
      關(guān)鍵詞:果皮

      劉鑫 張曉煜 孟君仁 段文宜 孫世航 潘磊 曾文芳 王志強(qiáng) 牛良

      摘? ? 要:【目的】探究桃果皮葉綠素降解規(guī)律及葉綠素降解相關(guān)基因的表達(dá)情況,以期為桃果實(shí)成熟期及底色差異判定確定依據(jù)?!痉椒ā恳源好?、2-50等不同桃品種/品系為研究對(duì)象,測(cè)量成熟前桃果實(shí)色差、葉綠素含量等指標(biāo),用熒光定量PCR檢測(cè)葉綠素降解相關(guān)基因的表達(dá)情況,通過(guò)分析葉綠素與相關(guān)基因表達(dá)的關(guān)系,確定影響桃果皮葉綠素降解的關(guān)鍵因素?!窘Y(jié)果】桃果實(shí)成熟前30 d葉綠素含量降低與PpCLH2、PpSGRL高表達(dá)有關(guān)。PpCLH1基因在桃果實(shí)轉(zhuǎn)色時(shí)表達(dá)量升高。在果實(shí)成熟前8~0 d,果皮葉綠素降解,PpSGR基因表達(dá)量上調(diào)。PpSGR基因表達(dá)情況與春美、2-50、中桃7號(hào)的葉綠素含量呈顯著負(fù)相關(guān),是導(dǎo)致桃果實(shí)成熟時(shí)葉綠素降解的關(guān)鍵基因?!窘Y(jié)論】成熟前12~8 d是桃果皮關(guān)鍵轉(zhuǎn)色期,PpSGR可能是桃果實(shí)成熟前果皮葉綠素降解的關(guān)鍵基因,這對(duì)進(jìn)一步解析桃果實(shí)發(fā)育過(guò)程中果皮葉綠素降解機(jī)制提供了一種新的思路,也為探索桃果實(shí)發(fā)育過(guò)程中葉綠素降解的分子機(jī)制提供參考。

      關(guān)鍵詞:桃;果皮;葉綠素降解;PpCLH1基因;PpSGR基因

      中圖分類號(hào):S662.1 文獻(xiàn)標(biāo)志碼:A 文章編號(hào):1009-9980(2024)06-1054-10

      Analysis of chlorophyll degradation and related gene expression in peach peel

      LIU Xin1, 2, ZHANG Xiaoyu1, 2, MENG Junren1, 2, DUAN Wenyi1, 2, SUN Shihang1, 2, PAN Lei1, 2, ZENG Wenfang1, 2, WANG Zhiqiang1, 2, NIU Liang1, 2*

      (1Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences/National Peach & Grape Improvement Center, Zhengzhou 450009, Henan, China; 2Zhongyuan Research Center, Chinese Academy of Agricultural Sciences, Xinxiang, 453004, Henan, China)

      Abstract: 【Objective】 Peach (Prunus persica), is one of the most important deciduous fruit trees in the world. The visual appeal of peach fruits, particularly their color, is a critical factor influencing consumer purchasing decisions. Chlorophyll content is important in determining the skin color and overall appearance of peaches. The precise role of chlorophyll degradation genes within the peach genome needs to fully understand, and their potential effects on chlorophyll breakdown in the peach peel and fruit ripening remain unclear. This study aimed to lay preliminary groundwork for understanding the ripening period and the underlying color differences in peach fruits by examining the color changes in the peach peels during the process of chlorophyll degradation, and the expression patterns of the genes related to the chlorophyll degradation. 【Methods】 Four peach varieties including Chunmei and 2-50 were chosen for this investigation. The color variations in the peach fruits prior to ripening were quantified using a colorimeter, and chlorophyll content was measured with a UV spectrophotometer. Although the primary chlorophyll degradation pathway has been identified, there are only limited researches on gene expression in the fruits. To fill this gap, quantitative transcription analysis was conducted, and the coding sequences (CDS) of relevant genes were sourced from genomic databases. The expression of the chlorophyll degradation-associated genes was quantified using real-time quantitative polymerase chain reaction (qRT-PCR), and the correlation between the chlorophyll degradation and the gene expression was established through statistical analysis. 【Results】 The L* value of the peach fruit color difference index increased gradually from 30 to 16 days before ripening and then decreased as the fruit matured. All the varieties exhibited an S-type a* value trend, with a* experiencing a sharp rise from 23 to 12 days prior to ripening. During the 12 days before ripening, the green hue of the peach fruit faded, and changed to red and showed a continuous increase in the a* value. The a* value shifted from negative to positive or near zero from 12 to 8 days before ripening, signifying the pivotal period for color change. The b* value initially increased from 30 to 16 days before ripening and then decreased, and the yellow-fleshed varieties showed higher values than the white-fleshed ones. The chlorophyll content significantly decreased in all the varieties as the fruit developed, and a steep decline was observed in 30 and 16 days before maturity. From 12 to 8 days before ripening, the chlorophyll levels of the four varieties fell below 8 ?g·g-1, indicating visible coloring, though ripening was still not completed. The high expression levels of the PpCLH2 and the PpSGRL were detected 30 days before maturity when the fruit was entirely green. The expression levels decreased as the fruit ripened, while the PpCLH1 expression peaked during the critical period of incomplete coloring and color transformation, suggesting that it would play a pivotal role in the chlorophyll degradation. The PpPPH, PpPAO and PpRCCR genes showed increased expression from 8 to 0 days before ripening, while the PpSGR was significantly overexpressed as the fruit matured, indicating that it would play a primary role in the chlorophyll degradation. The gene cluster analysis and correlation studies further highlighted the significant and negative correlation between the PpSGR expression and the chlorophyll content, underscoring its key function during peach fruit ripening. 【Conclusion】 This research would provide novel insights into the expression of the genes related to the chlorophyll degradation and their relationship with the color change and the chlorophyll content in the peach peels. We identified that the period approximately 12 to 8 days before ripening would be critical for the color transition in the peach peels, and the PpCLH1 and PpSGR would be the key genes in the pre-ripening chlorophyll degradation process. These findings would not only enhance our understanding of the chlorophyll degradation throughout peach fruit development but also offer a valuable reference for future studies on the molecular mechanisms of the chlorophyll degradation in peaches.

      Key words: Peach; Peel; Chlorophyll degradation; PpCLH1 gene; PpSGR gene

      隨著消費(fèi)者購(gòu)買(mǎi)力和消費(fèi)觀念的演變,果實(shí)大小、色澤、果面光潔度等外在品質(zhì)已成為影響消費(fèi)者購(gòu)買(mǎi)決策的主要因素[1],其中果皮底色對(duì)果實(shí)外觀有顯著影響,桃果實(shí)色澤和香味直接影響其感官品質(zhì),也是影響消費(fèi)者選擇的重要因素[2]。

      葉綠素是植物體的重要代謝產(chǎn)物[3],同樣也是決定果皮底色的關(guān)鍵因素。葉綠素降解是植物葉片衰老和果實(shí)成熟的一個(gè)重要代謝過(guò)程,葉綠素由多步驟途徑分解,其降解產(chǎn)生葉片變黃及果皮顏色變化,展現(xiàn)果實(shí)成熟的狀態(tài)[4],所以全面、深入地了解葉綠素降解及其關(guān)鍵基因的調(diào)控機(jī)制,可以加深對(duì)桃果實(shí)成熟時(shí)色澤變化的理解。近年來(lái),植物中葉綠素的主要降解途徑已有較多的研究[5-6]。葉綠素降解途徑中葉綠素b首先被NYC1和NOL兩種同工酶還原生成7-羥甲基葉綠素a;隨后,7-羥甲基葉綠素a被HCAR還原為葉綠素a[7];進(jìn)而,葉綠素a被葉綠素酶(CLH)催化形成脫植基葉綠素[8],在金屬螯合物作用下先除去鎂離子生成脫鎂葉綠素,生成的Pheina則在脫鎂葉綠素酶(PPH)作用下除去植基[9];之后PAO能將脫鎂基葉綠酸a(Pheidea)降解為不穩(wěn)定的紅色葉綠素代謝產(chǎn)物(RCC)后,最終在RCCR的催化下形成無(wú)色的初生熒光葉綠素代謝產(chǎn)物(pFCC)[10]。

      PAO是控制葉綠素降解的重要基因[11-12],PAO催化的卟啉環(huán)氧化開(kāi)環(huán)是葉綠素降解的關(guān)鍵步驟,因而這條降解途徑被稱為PAO降解途徑。棗中ZjPAO1和ZjPAO2基因在果實(shí)不同發(fā)育階段均有表達(dá),并會(huì)隨著果實(shí)發(fā)育表達(dá)量升高[13]。水稻中發(fā)現(xiàn)一個(gè)單堿基突變導(dǎo)致PAO編碼蛋白變化,從而導(dǎo)致水稻中活性氧積累和葉綠素降解受阻[14]。SGR和SGRL基因是葉綠素降解調(diào)控途徑中具有里程碑意義的基因。SGR與SGRL基因與葉綠素降解相關(guān),可以通過(guò)招募葉綠素降解基因形成復(fù)合體,結(jié)合到光系統(tǒng)Ⅱ上,從而導(dǎo)致葉綠素降解。但相比于SGR,SGRL在植物衰老前表達(dá)量較高,隨后表達(dá)量開(kāi)始下降[15]。枳砧紅綿蜜柚嫁接黃化苗中凈光合速率、葉綠素含量降低,SGR表達(dá)量升高[16]。在小麥中發(fā)現(xiàn)在自然衰老7 d時(shí)CLH表達(dá)量升高,之后下降[17]。在大白菜中,MeJA處理顯著誘導(dǎo)葉綠素降解相關(guān)基因BrPAO1、BrNYC1、BrPPH1和BrSGR1的表達(dá)[18]。在柑橘中發(fā)現(xiàn),宗橙中CsSGRaSTOP喪失了降解葉綠素的功能,導(dǎo)致其果皮呈棕褐色[19]。在香蕉中發(fā)現(xiàn)MaERF012在果肉和果皮中差異表達(dá),并與果實(shí)成熟密切相關(guān),MaERF012激活葉綠素降解基因MaSGR的表達(dá)從而導(dǎo)致香蕉果皮葉綠素降解[20]。擬南芥在葉片自然衰老過(guò)程中,CCGs劇烈上調(diào)表達(dá)且CCEs蛋白逐漸累積,導(dǎo)致衰老細(xì)胞中葉綠素的大量降解[21]。

      桃果皮色澤是影響食用價(jià)值和經(jīng)濟(jì)價(jià)值的重要因素之一,葉綠素降解及相關(guān)基因調(diào)控在部分物種中有一定的研究,但在桃中葉綠素降解及相關(guān)基因的表達(dá)規(guī)律尚未見(jiàn)報(bào)道[22]。筆者在本研究中通過(guò)熒光定量技術(shù)對(duì)4個(gè)桃品種/品系果皮葉綠素降解基因表達(dá)規(guī)律進(jìn)行研究,并系統(tǒng)分析了葉綠素降解相關(guān)基因表達(dá)規(guī)律與葉綠素水平和色差之間的關(guān)系。

      1 材料和方法

      1.1 試驗(yàn)材料

      以4個(gè)不同成熟期的桃品種/品系春美、2-50、中蟠102、中桃7號(hào)為研究材料(圖1),其中,春美和2-50在6月中旬成熟,中蟠102和中桃7號(hào)分別在7月上旬和8月上旬成熟。供試材料來(lái)自中國(guó)農(nóng)業(yè)科學(xué)院鄭州果樹(shù)研究所桃品種圃,株行距為1.0 m×4.0 m,2017年定植,常規(guī)管理。2022年5—8月,于果實(shí)成熟前30 d開(kāi)始,采摘樹(shù)體外圍中上部大小均勻、無(wú)病蟲(chóng)害、成熟度一致的果實(shí)30個(gè),帶回實(shí)驗(yàn)室使用尼康700d相機(jī)于自然光下以黑色植絨布為背景拍照。以10個(gè)果實(shí)為1個(gè)樣本,取樣進(jìn)行3次重復(fù),取樣間隔7 d,在果實(shí)轉(zhuǎn)色期每隔4 d取樣1次,削取表皮進(jìn)行液氮速凍,保存至-80 ℃冰箱備用。

      1.2 色差檢測(cè)

      用色差儀(美能達(dá)CR-400,柯尼卡美能達(dá))評(píng)價(jià)果皮底色,顏色用CIE a*、b*標(biāo)尺表示。隨機(jī)選取果實(shí)赤道區(qū)域的四個(gè)不同點(diǎn)取平均值,記錄a*和b*值[23-24]。

      1.3 葉綠素含量測(cè)定

      桃果皮葉綠素含量采用紫外分光光度法測(cè)定[25-26]。取去離子水清洗過(guò)的桃果皮0.4 g,用研磨機(jī)磨碎,放入10 mL離心管中,加入6 mL 95%乙醇溶液,避光提取葉綠素至果皮完全變白。分別測(cè)定葉綠素溶液在665 nm和649 nm波長(zhǎng)下的吸光度,通過(guò)公式1~3計(jì)算葉綠素含量(w,后同):

      1.4 RNA提取

      根據(jù)多糖多酚植物總RNA提取試劑盒(DP441,天根生化科技有限公司,北京,中國(guó))說(shuō)明書(shū)提取桃總RNA。利用1%瓊脂糖凝膠電泳檢測(cè)RNA的質(zhì)量和純度,取1 ?L RNA利用微量紫外分光光度計(jì)NanoDrop2000(Thermo Scientific,麻?。y(cè)定濃度。取1 ng RNA參照FastKing cDNA第一鏈合成試劑盒說(shuō)明書(shū)(天根生化科技有限公司,北京,中國(guó))進(jìn)行反轉(zhuǎn)錄,放置在-20 ℃冰箱保存進(jìn)行后續(xù)的實(shí)時(shí)熒光定量PCR。

      1.5 實(shí)時(shí)熒光定量檢測(cè)

      從基因組數(shù)據(jù)庫(kù)(https://phytozome-next.jgi.doe.gov/)下載相關(guān)基因的CDS(Coding sequence,編碼序列);利用NCBI網(wǎng)站(https://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?LINK_LOC=BlastHome)設(shè)計(jì)特異性熒光定量引物,引物長(zhǎng)度在20 bp左右,GC含量為40%~60%,引物Tm值為58 ℃~62 ℃,擴(kuò)增片段大小為80~150 bp;PCR體系為15.1 ?L:cDNA 1 ?L、Mix 7.5 ?L、引物F 0.3 ?L、引物R 0.3 ?L、水6 ?L。PCR程序設(shè)置為:95 ℃預(yù)變性5 min;95 ℃變性30 s、60 ℃退火30 s、72 ℃延伸30 s,擴(kuò)增循環(huán)數(shù)為40,設(shè)置3次技術(shù)重復(fù)。引物序列見(jiàn)表1。實(shí)時(shí)熒光定量PCR所用的儀器為羅氏480,所用試劑盒為SYBR Real-time PCR Premixtur(eBiotake)。選取Actin(ppa007228mg)為桃內(nèi)參基因[27],最后按照2-ΔΔCT方法進(jìn)行結(jié)果計(jì)算[28]。

      1.6 數(shù)據(jù)分析

      試驗(yàn)數(shù)據(jù)用SPSS軟件分析,用Excel 2003軟件制作圖表。

      2 結(jié)果與分析

      2.1 桃果實(shí)發(fā)育后期果皮色差變化規(guī)律

      2.1.1 a*值的變化 4個(gè)品種/品系a*值(圖2)均呈現(xiàn)為S形,在成熟前23~12 d色差a*上升較快。在果實(shí)成熟前12 d,桃果實(shí)綠色不斷減弱,果實(shí)開(kāi)始著紅色,色差a*值不斷上升。在成熟前12~8 d,4個(gè)品種果實(shí)色差a*值均由負(fù)轉(zhuǎn)正或接近于0,認(rèn)為此時(shí)是果實(shí)重要的轉(zhuǎn)色期。

      2.1.2 b*值的變化 中蟠102和中桃7號(hào)果實(shí)b*值(圖3)在成熟前30~16 d升高,之后隨著果實(shí)發(fā)育不斷下降。春美和2-50果實(shí)b*值在成熟前30~23 d升高,隨后快速下降。春美、2-50、中桃7號(hào)在果實(shí)成熟時(shí)b*值為20~30。

      2.2 果實(shí)發(fā)育后期葉綠素含量變化

      4個(gè)品種/品系的葉綠素含量(圖4)隨著果實(shí)的發(fā)育表現(xiàn)出明顯的下降趨勢(shì),葉綠素含量在成熟前30~16 d快速下降。在果實(shí)成熟前12~8 d,各品種的葉綠素含量(w)均在8 μg·g-1以下,從圖1可以看到這時(shí)各品種有明顯著色,但果實(shí)著色還不完全。在果實(shí)成熟前8 d至果實(shí)成熟,4個(gè)桃品種/品系的葉綠素含量下降較為緩慢。中蟠102和2-50在成熟前8 d時(shí)葉綠素含量與成熟時(shí)的葉綠素含量差異不大。

      2.3 果實(shí)發(fā)育后期葉綠素降解相關(guān)基因的表達(dá)分析

      通過(guò)實(shí)時(shí)熒光定量PCR方法,測(cè)定了4個(gè)桃品種/品系中與葉綠素降解相關(guān)的10個(gè)基因(PpNYC1、PpNOL、PpHCAR、PpCLH1、PpCLH2、PpPPH、PpPAO、PpRCCR、PpSGR、PpSGRL)的表達(dá)量,并繪制熱圖進(jìn)行聚類分析。如圖5所示,PpCLH1、PpPPH、PpPAO、PpRCCR、PpSGR、PpSGRL基因在春美材料中高表達(dá),PpNYC1在中蟠102中的表達(dá)量較高,PpNOL在中桃7號(hào)中表達(dá)量最高,PpCLH2則在中蟠102和中桃7號(hào)高表達(dá),而PpHCAR在4個(gè)桃品種/品系中無(wú)明顯表達(dá)差異。

      不同時(shí)間段內(nèi),每個(gè)基因的表達(dá)量通常會(huì)隨著果實(shí)發(fā)育不斷變化。PpNYC1在中蟠102中成熟前4 d時(shí)有升高,PpNOL在中桃7號(hào)中明顯先上調(diào)后下調(diào),PpSGRL在中桃7號(hào)成熟前12 d有短暫升高。PpHCAR、PpPPH、PpPAO、PpRCCR隨著果實(shí)發(fā)育在成熟前12~4 d表達(dá)量上升,在果實(shí)成熟時(shí)表達(dá)量下降,PpHCAR、PpRCCR在3個(gè)桃品種/品系中無(wú)明顯差異,在2-50中表達(dá)模式有所不同。PpPPH和PpPAO在春美中分別在成熟前8 d和成熟前4 d有明顯升高。

      由圖5可知,在4個(gè)桃品種/品系中,PpCLH1基因在成熟前12~8 d時(shí)間段內(nèi)的表達(dá)量高于其他基因,在果實(shí)成熟時(shí)表達(dá)量迅速下降,因?yàn)榇藭r(shí)果實(shí)處于轉(zhuǎn)色的關(guān)鍵期,因此推斷PpCLH1基因可能是桃果實(shí)轉(zhuǎn)色時(shí)的重要調(diào)控基因。隨著果實(shí)發(fā)育,果皮葉綠素降解,果實(shí)完全成熟,PpSGR基因的表達(dá)量顯著升高。進(jìn)一步推測(cè)PpSGR基因有可能也是桃果實(shí)成熟前造成4個(gè)桃品種/品系葉綠素降解的主要基因。

      根據(jù)聚類分析結(jié)果(圖6)在春美和2-50中PpSGR與PpCLH1、PpPPH聚為一類,在中蟠102和中桃7號(hào)中PpSGR與PpCLH1、PpRCCR聚為一類。在春美和中蟠102中PpSGRL和PpCLH2聚為一類,在2-50和中桃7號(hào)中PpSGRL、PpNYC1和PpCLH2聚為一類。因此PpSGRL、PpNYC1和PpCLH2可能是果實(shí)早期葉綠素降解的關(guān)鍵基因,PpSGR與PpCLH1、PpPPH、PpRCCR可能是桃果實(shí)成熟前造成不同桃品種/品系葉綠素降解差異的主要基因。

      對(duì)葉綠素含量與其降解基因進(jìn)行相關(guān)性分析(表2),發(fā)現(xiàn)PpCLH2基因表達(dá)與春美和2-50中葉綠素含量呈顯著正相關(guān),PpSGR與春美、中桃7號(hào)的葉綠素含量呈顯著負(fù)相關(guān),PpSGRL與春美、2-50、中蟠102的葉綠素含量呈顯著正相關(guān)。

      qRT-PCR分析結(jié)果表明,成熟前12~8 d時(shí),PpCLH1、PpPAO基因的表達(dá)量顯著。而PpSGR基因在果實(shí)成熟時(shí)表達(dá)量明顯上調(diào),聚類分析顯示在春美和2-50中PpSGR與PpCLH1、PpPPH聚為一類,在中蟠102和中桃7號(hào)中PpSGR與PpCLH1、PpRCCR聚為一類。相關(guān)性分析顯示PpSGR與春美、中桃7號(hào)的葉綠素含量呈顯著負(fù)相關(guān)。初步表明PpCLH1、PpSGR基因是果實(shí)成熟前4個(gè)桃品種/品系果皮葉綠素降解的關(guān)鍵基因。

      3 討 論

      長(zhǎng)期以來(lái),CLH被認(rèn)為是參與葉綠素降解的第一種酶,與葉綠素的降解密切相關(guān)。有研究表明,乙烯處理的柑橘果皮中CLH酶活力提高了2.5~4倍,同時(shí)果皮明顯褪綠[29]。樊艷燕等[30]發(fā)現(xiàn)BoCLH1(葉綠素酶1)在青花菜初始時(shí)檢測(cè)到有較高表達(dá),后期表達(dá)量很低,試驗(yàn)結(jié)果表明BoCLH1主要在青花菜初始衰老中起降解葉綠素的作用。胡椒中發(fā)現(xiàn)葉綠素酶活性與葉綠素含量之間存在負(fù)相關(guān)關(guān)系,并在收獲后的衰老過(guò)程中促進(jìn)葉綠素的降解[31]。筆者在本試驗(yàn)中發(fā)現(xiàn),桃PpCLH1在果實(shí)成熟過(guò)程中表達(dá)量先升高后下降,在果實(shí)成熟前12~8 d表達(dá)量較高,此時(shí)也是果實(shí)轉(zhuǎn)色的關(guān)鍵時(shí)期,因此PpCLH1可能是桃果實(shí)轉(zhuǎn)色時(shí)葉綠素降解的關(guān)鍵基因。在草地早熟禾葉片衰老時(shí),CLH基因的表達(dá)會(huì)導(dǎo)致葉綠素降解[32]。在荔枝中成熟過(guò)程中LcCLH表達(dá)量先升高后下降,LcPAO、LcSGR的表達(dá)量隨著果實(shí)發(fā)育逐漸升高,相比于葉綠素降解更徹底的糯米滋,妃子笑中LcCLH、LcPAO、LcSGR的表達(dá)量明顯更低[33]。

      在高等植物中,脫鎂葉綠酸a氧化酶(PAO)基因表達(dá)量和酶活性的變化與葉綠素含量降解有關(guān)[12]。此外SGR普遍參與了植物葉片衰老過(guò)程中葉綠素分解,也在植物其他器官的生長(zhǎng)發(fā)育中有重要作用,如果實(shí)成熟和發(fā)育[34]。在大豆成熟前PPH、PAO、SGR三個(gè)基因相對(duì)表達(dá)量顯著提高(p<0.05),并且在褪綠材料科豐14中的表達(dá)量遠(yuǎn)高于滯綠材料北農(nóng)108中的表達(dá)量,因此推測(cè)葉綠素降解基因PPH、PAO、SGR表達(dá)量低是北農(nóng)108滯綠的重要原因之一[35]。在本試驗(yàn)中PpPPH和PpPAO在春美中分別在8 DBM和4 DBM有明顯升高,在其他4個(gè)品種/品系中也都有明顯升高。雷波臍橙及其早熟單株果實(shí)也發(fā)現(xiàn)PAO在果實(shí)各發(fā)育時(shí)期都有穩(wěn)定表達(dá),CLH、PPH、NYC1、SGR1的表達(dá)先上調(diào)后下調(diào)[36]。青甌柑和甌柑的試驗(yàn)結(jié)果表明在花后210 d、240 d,甌柑中NYC1、CLH、PPH、PAO、SGR的表達(dá)量均高于青甌柑,差異顯著,這可能是青甌柑葉綠素降解緩慢的原因[37]。筆者在本研究中通過(guò)聚類分析,在春美和2-50中PpSGR與PpCLH1、PpPPH聚為一類,在中蟠102和中桃7號(hào)中PpSGR與PpCLH1、PpRCCR聚為一類。這說(shuō)明在桃中葉綠素降解受多個(gè)葉綠素降解基因的影響。

      大豆中GmSGR丟失可能導(dǎo)致大豆種子從黃色變?yōu)榫G色[38]。擬南芥在葉片衰老過(guò)程中,SGR1正向介導(dǎo)葉綠素降解,而過(guò)表達(dá)SGRL的擬南芥植物表現(xiàn)出早期葉片黃變[15]。葉綠素降解導(dǎo)致桃果皮褪綠是桃果實(shí)成熟最明顯的標(biāo)志之一。有研究表明SlSGR1在調(diào)節(jié)番茄葉片和果實(shí)的葉綠素降解中起著關(guān)鍵作用[39]。與野生型(WT)果實(shí)相比,SlSGR1敲除無(wú)效系明顯顯示出混濁的棕色,葉綠素水平明顯更高[17],表明SlSGR1影響葉綠素降解。筆者課題組發(fā)現(xiàn)PpSGR基因在果實(shí)成熟時(shí)表達(dá)量較高,并且春美、中桃7號(hào)葉綠素含量與PpSGR表達(dá)量呈負(fù)相關(guān),所以PpSGR是桃果實(shí)成熟期葉綠素降解的關(guān)鍵調(diào)控基因。目前在其他物種中葉綠素降解基因的表達(dá)規(guī)律與功能的研究已經(jīng)有很多,但在桃中還少見(jiàn)報(bào)道。因此,進(jìn)一步解析桃中葉綠素降解基因調(diào)控果實(shí)成熟前葉綠素降解的機(jī)制具有重要意義。

      4 結(jié) 論

      桃果實(shí)成熟前12~8 d是桃果實(shí)關(guān)鍵的轉(zhuǎn)色期,此時(shí)PpCLH1基因相比綠色時(shí)有明顯上調(diào)。在果實(shí)成熟前8~0 d,隨著果皮葉綠素降解,PpSGR表達(dá)量上升,表明PpSGR是桃果實(shí)成熟前葉綠素降解的關(guān)鍵基因。本研究為探索桃果實(shí)發(fā)育過(guò)程中葉綠素降解的分子機(jī)制提供了參考。

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      收稿日期:2024-01-04 接受日期:2024-03-25

      基金項(xiàng)目:國(guó)家桃產(chǎn)業(yè)技術(shù)體系(CARS-30);中國(guó)農(nóng)業(yè)科學(xué)院科技創(chuàng)新工程專項(xiàng)經(jīng)費(fèi)項(xiàng)目(CAAS-ASTIP-2023-ZFRI);河南省重大公益科技專項(xiàng)(201300110500)

      作者簡(jiǎn)介:劉鑫,男,在讀碩士研究生,研究方向?yàn)楣麡?shù)遺傳育種。E-mail:82101212241@caas.cn

      *通信作者 Author for correspondence. E-mail:niuliang@caas.cn

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