文冠果果实韧皮部及其周围薄壁细胞的超微结构观察及功能分析
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摘要
该研究应用透射电镜技术,对生长发育过程中的文冠果果实的韧皮部及其周围薄壁细胞的超微结构进行了观察,以探讨文冠果果实同化物韧皮部卸载的细胞学路径及其机理。结果显示:(1)文冠果果实发育过程中,筛分子细胞胞腔较空,几乎没有细胞器,但有类似于囊泡的丝状不定型物存在;伴胞胞质浓密且细胞器丰富,液泡化程度不一,大多数存在多个小液泡;薄壁细胞具有中央大液泡,发育中期富含线粒体、高尔基体、内质网等细胞器,并存在囊泡运输现象,发育后期细胞器发生降解,说明随着果实生长发育,果实内物质代谢和转运活跃程度逐渐下降。(2)果实发育过程中筛分子和伴胞之间始终有胞间连丝,薄壁细胞之间也一直存在大量的胞间连丝,而筛分子-伴胞复合体与薄壁细胞之间只有在果实发育前期和后期存在一定数量的胞间连丝,发育中期却几乎没有胞间连丝。研究结果表明,文冠果果实发育过程中同化物韧皮部卸载路径可能发生了共质体途径-质外体途径-共质体途径的转变。
In this study, we systematically investigated the ultrastructure of the phloem and surrounding parenchyma cells using transmission electron microscopy during the development process of Xanthoceras sorbifolium Bunge fruit, to understand the unloading pathway and mechanism during the fruit development. The results showed:(1) during the fruit development process, the cell cavity of the sieve element(SE)was empty and few organelles exited, but there were some vesicle-like and microfibril-like structures in SE. With increase of cytoplasm density and organelles abundance, the degree of vacuolization in companion cell(CC)varied, and most of them had multiple small vacuoles; Phloem parenchyma cell(PP)had large central vacuoles, and vesicle transport occurred in the middle stage of development. They were rich in organelles such as mitochondria, golgi bodies and endoplasmic reticulum, which degraded at the later stage of development. This indicated that the metabolism and substances transport in fruit decreased gradually with the fruit development.(2) During the development of fruit, there were always some plasmodesmata between SE and CC, and lots of plasmodesmata between CC. However, only a certain number of plasmodesmata existed between the SE-CC complex and PP at the early and late stage of fruit development, but almost no plasmodesmata existed at the middle stage of fruit development. Based on the above results, we concluded that the unloading pathway of assimilate changed from symplastic to apoplastic and then to symplastic during the fruit development.
In this study, we systematically investigated the ultrastructure of the phloem and surrounding parenchyma cells using transmission electron microscopy during the development process of Xanthoceras sorbifolium Bunge fruit, to understand the unloading pathway and mechanism during the fruit development. The results showed:(1) during the fruit development process, the cell cavity of the sieve element(SE)was empty and few organelles exited, but there were some vesicle-like and microfibril-like structures in SE. With increase of cytoplasm density and organelles abundance, the degree of vacuolization in companion cell(CC)varied, and most of them had multiple small vacuoles; Phloem parenchyma cell(PP)had large central vacuoles, and vesicle transport occurred in the middle stage of development. They were rich in organelles such as mitochondria, golgi bodies and endoplasmic reticulum, which degraded at the later stage of development. This indicated that the metabolism and substances transport in fruit decreased gradually with the fruit development.(2) During the development of fruit, there were always some plasmodesmata between SE and CC, and lots of plasmodesmata between CC. However, only a certain number of plasmodesmata existed between the SE-CC complex and PP at the early and late stage of fruit development, but almost no plasmodesmata existed at the middle stage of fruit development. Based on the above results, we concluded that the unloading pathway of assimilate changed from symplastic to apoplastic and then to symplastic during the fruit development.
引文
[1] OPARKA K J.What is phloem unloading?[J].Plant Physiology,1990,94(2):393-396.
[2] PATRICK J W.PHLOEM UNLOADING:Sieve element unloading and post-sieve element transport[J].Plant Biology,1997,48(48):191-222.
[3] ZHANG L Y,PENG Y B,et al.Evidence for apoplasmic phloem unloading in developing apple fruit [J].Plant Physiology,2004,135(1):574-586.
[4] WU G L,ZHANG X Y,ZHANG L Y,et al.Phloem unloading in developing walnut fruit is symplasmic in the seed pericarp and apoplasmic in the fleshy pericarp [J].Plant Cell Physiology,2004,45(10):1 461-1 470.
[5] ZHANG X Y,WANG X L,et al.A shift of phloem unloading from symplasmic to apoplasmic pathway is involved in developmental onset of ripening in grape berry [J].Plant Physiology,2006,142(1):220-232.
[6] ZHANG H P,WU J Y,TAO S T,et al.Evidence for apoplasmic phloem unloading in pear fruit[J].Plant Molecular Biology Reporter,2014,32(4):931-939.
[7] ZANON L,FALCHI R,et al.Sucrose transport and phloem unloading in peach fruit:potential role of two transporters localized in different cell types[J].Physiologia Plantarum,2015,154(2):179-193.
[8] WANG T D,ZHANG H F,WU Z C,et al.Sugar uptake in the aril of litchi fruit depends on the apoplasmic post-phloem transport and the activity of proton pumps and the putative transporter LcSUT4[J].Plant and Cell Physiology,2015,56(2):377-387.
[9] NIE P,WANG X,HU L,et al.The predominance of the apoplasmic phloem-unloading pathway is interrupted by a symplasmic pathway during Chinese jujube fruit development [J].Plant Cell Physiology,2010,51(6):1 007-1 018.
[10] 张鹤华,李艳芳,聂佩显,等.蓝莓果实同化物韧皮部卸载路径与糖代谢酶活性[J].林业科学,2017,53(3):40-48.ZHANG H H,LI Y F,NIE P X,et al.Phloem unloading pathway of photosynthates and sucrose-metabolizing enzymes activities in Vaccinium corymbosum fruit[J].Scientia Silvae Sinicae,2017,53(3):40-48.
[11] 李春丽.无花果果实生理指标及其韧皮部卸载途径研究[D].乌鲁木齐:新疆农业大学,2016.
[12] 郑国琦,包晗,杨涓,等.宁夏枸杞果实韧皮部及其周围细胞超微结构研究[J].西北植物学报,2015,35(11):2 211-2 218.ZHENG G Q,BAO H,YANG J,et al.Ultrastructure of phloem and the flesh sink-cells during fruit development of Lycium barbarum[J].Acta Botanica Boreali-Occidentalia Sinica,2015,35(11):2 211-2 218.
[13] HU L,SUN H,LI R,et al.Phloem unloading follows an extensive apoplasmic pathway in cucumber (Cucumis sativus L.) fruit from anthesis to marketable maturing stage[J].Plant Cell & Environment,2011,34(11):1 835-1 848.
[14] CHEN C,YUAN Y,ZHANG C,et al.Sucrose phloem unloading follows an apoplastic pathway with high sucrose synthase in actinidia fruit[J].Plant Science,2017,255:40-50.
[15] WANG X Y,YOU H,et al.The cellular pathway and enzymatic activity for phloem-unloading transition in developing Camellia oleifera Abel.fruit[J].Acta Physiologiae Plantarum,2018,40(2):23.
[16] 岳胜钱,栗燕,杨秋生.植物光合同化物韧皮部卸载途径研究进展[J].河南农业科学,2016,45(4):1-6.YUE S Q,LI Y,YANG Q S.Research progress on phloem unloading pathway of plant photoassimilate[J].Journal of Henan Agricultural Sciences,2016,45(4):1-6.
[17] 聂佩显,关秋竹,张振贤,等.冬枣果实韧皮部及其周围薄壁细胞的超微结构观察和功能分析[J].电子显微学报,2009,28(4):389-395.NIE P X,GUAN Q Z,ZHANG Z X,et al.Ultrastructures of phloem and its surrounding parenchyma cells in the developing Zizyphus jujuba Mill.cv Dongzao fruit[J].Journal of Chinese Electron Microscopy Society,2009,28(4):389-395.
[18] 敖妍,马履一.文冠果分子生物学研究进展[J].分子植物育种,2016,14(12):3 570-3 576.AO Y,MA L Y.Research progress in molecular biology of Xanthoceras sorbifolia Bunge[J].Molecular Plant Breeding,2016,14(12):3 570-3 576.
[19] 高述民,马凯,杜希华,等.文冠果(Xanthoceras sorbifolia)研究进展[J].植物学通报,2002,19(3):296-301.GAO S M,MA K,DU X H,et al.Advances in research on Xanthoceras sorbifolia[J].Chinese Bulletin of Botany,2002,19(3):296-301.
[20] 敖妍,段劼,于海燕,等.文冠果研究进展[J].中国农业大学学报,2012,17(6):197-203.AO Y,DUAN J,YU H Y,et al.Research progress on Xanthoceras sorbifolia Bunge[J].Journal of China Agricultural University,2012,17(6):197-203.
[21] 苏曼琳,吴尚,马履一,等.3种修剪方法对文冠果芽内源激素含量和萌芽成枝的影响[J].西北农林科技大学学报(自然科学版),2017,45(5):101-108.SU M L,WU S,MA L Y,et al.Effects of three pruning methods on endogenous hormone and shoot growth of Xanthoceras sorbifolia Bunge buds[J].Journal of Northwest A&F University (Natural Science Edition),2017,45(5):101-108.
[22] 史绍林.文冠果种子催芽和组织培养研究[D].黑龙江齐齐哈尔:齐齐哈尔大学,2012.
[23] 郝一男,王喜明,丁立军.文冠果油生物柴油副产物粗甘油的精制[J].中国油脂,2011,36(11):80-82.HAO Y N,WANG X M,DING L J.Refining of crude glycerol produced from the production of Xanthoceras sorbiflia Bunge seed oil biodiesel[J].China Oils and Fats,2011,36(11):80-82.
[24] 孙俊.文冠果油的提取及其生物柴油制备工艺研究[D].西安:陕西师范大学,2008.
[25] 白鑫磊,金华,邹吉祥,等.文冠果表型性状的相关性及主成分分析[J].经济林研究,2019,37(1):68-73.BAI X L,JIN H,ZOU J X,et al.Correlation analysis and principal component analysis on phenotypic characteristics in Xanthoceras sorbifolia[J].Nonwood Forest Research,2019,37(1):68-73.
[26] 马新,姜继元,李铭.24个种源文冠果在新疆的生长结实性状分析[J].经济林研究,2019,37(1):61-67.MA X,JIANG J Y,LI M.Analysis on growth and fruiting characteristics of Xanthoceras sorbifolia from 24 provenances in Xinjiang[J].Nonwood Forest Research,2019,37(1):61-67.
[27] 路蒙蒙,韩硕,杨琦,等.文冠果LEC1基因的克隆及表达分析[J].北京林业大学学报,2018,40(1):8-16.LU M M,HAN S,YANG Q,et al.Cloning and expression analysis of LEC1 gene from Xanthoceras sorbifolia embryos[J].Journal of Beijing Forestry University,2018,40(1):8-16.
[28] 赵娜,张媛,李秋琦,等.文冠果FAD2的序列与功能分析[J].北京林业大学学报,2015,37(2):87-93.ZHAO N,ZHANG Y,LI Q Q,et al.Sequence and functional analysis of FAD2 gene from Xanthoceras sorbifolia seeds[J].Journal of Beijing Forestry University,2015,37(2):87-93.
[29] 刘祾悦,阮成江,王莉,等.文冠果种仁油脂脂肪酸形成的多基因协同调控[J].分子植物育种,2019,17(6):1 834-1 842.LIU L Y,RUAN C J,WANG L,et al.Coordinated regulation of multigenes formed by fatty acids in kernel oil of Xanthoceras sorbifolium[J].Molecular Plant Breeding,2019,17(6):1 834-1 842.
[30] 赵翠格.文冠果种子发育过程中油脂累积规律研究[D].北京:北京林业大学,2010.
[31] 张宏平,张晋元,刘玉香,等.桃果实组织结构和超微结构细胞学研究[J].北方园艺,2018,(1):29-34.ZHANG H P,ZHANG J Y,LIU Y X,et al.Histological and ultrastructural cytology of peach fruit[J].Northern Horticulture,2018,(1):29-34.
[32] RUAN Y L,PATRICK J W.The cellular pathway of postphloem sugar transport in developing tomato fruit[J].Planta,1995,196(3):434-444.
[2] PATRICK J W.PHLOEM UNLOADING:Sieve element unloading and post-sieve element transport[J].Plant Biology,1997,48(48):191-222.
[3] ZHANG L Y,PENG Y B,et al.Evidence for apoplasmic phloem unloading in developing apple fruit [J].Plant Physiology,2004,135(1):574-586.
[4] WU G L,ZHANG X Y,ZHANG L Y,et al.Phloem unloading in developing walnut fruit is symplasmic in the seed pericarp and apoplasmic in the fleshy pericarp [J].Plant Cell Physiology,2004,45(10):1 461-1 470.
[5] ZHANG X Y,WANG X L,et al.A shift of phloem unloading from symplasmic to apoplasmic pathway is involved in developmental onset of ripening in grape berry [J].Plant Physiology,2006,142(1):220-232.
[6] ZHANG H P,WU J Y,TAO S T,et al.Evidence for apoplasmic phloem unloading in pear fruit[J].Plant Molecular Biology Reporter,2014,32(4):931-939.
[7] ZANON L,FALCHI R,et al.Sucrose transport and phloem unloading in peach fruit:potential role of two transporters localized in different cell types[J].Physiologia Plantarum,2015,154(2):179-193.
[8] WANG T D,ZHANG H F,WU Z C,et al.Sugar uptake in the aril of litchi fruit depends on the apoplasmic post-phloem transport and the activity of proton pumps and the putative transporter LcSUT4[J].Plant and Cell Physiology,2015,56(2):377-387.
[9] NIE P,WANG X,HU L,et al.The predominance of the apoplasmic phloem-unloading pathway is interrupted by a symplasmic pathway during Chinese jujube fruit development [J].Plant Cell Physiology,2010,51(6):1 007-1 018.
[10] 张鹤华,李艳芳,聂佩显,等.蓝莓果实同化物韧皮部卸载路径与糖代谢酶活性[J].林业科学,2017,53(3):40-48.ZHANG H H,LI Y F,NIE P X,et al.Phloem unloading pathway of photosynthates and sucrose-metabolizing enzymes activities in Vaccinium corymbosum fruit[J].Scientia Silvae Sinicae,2017,53(3):40-48.
[11] 李春丽.无花果果实生理指标及其韧皮部卸载途径研究[D].乌鲁木齐:新疆农业大学,2016.
[12] 郑国琦,包晗,杨涓,等.宁夏枸杞果实韧皮部及其周围细胞超微结构研究[J].西北植物学报,2015,35(11):2 211-2 218.ZHENG G Q,BAO H,YANG J,et al.Ultrastructure of phloem and the flesh sink-cells during fruit development of Lycium barbarum[J].Acta Botanica Boreali-Occidentalia Sinica,2015,35(11):2 211-2 218.
[13] HU L,SUN H,LI R,et al.Phloem unloading follows an extensive apoplasmic pathway in cucumber (Cucumis sativus L.) fruit from anthesis to marketable maturing stage[J].Plant Cell & Environment,2011,34(11):1 835-1 848.
[14] CHEN C,YUAN Y,ZHANG C,et al.Sucrose phloem unloading follows an apoplastic pathway with high sucrose synthase in actinidia fruit[J].Plant Science,2017,255:40-50.
[15] WANG X Y,YOU H,et al.The cellular pathway and enzymatic activity for phloem-unloading transition in developing Camellia oleifera Abel.fruit[J].Acta Physiologiae Plantarum,2018,40(2):23.
[16] 岳胜钱,栗燕,杨秋生.植物光合同化物韧皮部卸载途径研究进展[J].河南农业科学,2016,45(4):1-6.YUE S Q,LI Y,YANG Q S.Research progress on phloem unloading pathway of plant photoassimilate[J].Journal of Henan Agricultural Sciences,2016,45(4):1-6.
[17] 聂佩显,关秋竹,张振贤,等.冬枣果实韧皮部及其周围薄壁细胞的超微结构观察和功能分析[J].电子显微学报,2009,28(4):389-395.NIE P X,GUAN Q Z,ZHANG Z X,et al.Ultrastructures of phloem and its surrounding parenchyma cells in the developing Zizyphus jujuba Mill.cv Dongzao fruit[J].Journal of Chinese Electron Microscopy Society,2009,28(4):389-395.
[18] 敖妍,马履一.文冠果分子生物学研究进展[J].分子植物育种,2016,14(12):3 570-3 576.AO Y,MA L Y.Research progress in molecular biology of Xanthoceras sorbifolia Bunge[J].Molecular Plant Breeding,2016,14(12):3 570-3 576.
[19] 高述民,马凯,杜希华,等.文冠果(Xanthoceras sorbifolia)研究进展[J].植物学通报,2002,19(3):296-301.GAO S M,MA K,DU X H,et al.Advances in research on Xanthoceras sorbifolia[J].Chinese Bulletin of Botany,2002,19(3):296-301.
[20] 敖妍,段劼,于海燕,等.文冠果研究进展[J].中国农业大学学报,2012,17(6):197-203.AO Y,DUAN J,YU H Y,et al.Research progress on Xanthoceras sorbifolia Bunge[J].Journal of China Agricultural University,2012,17(6):197-203.
[21] 苏曼琳,吴尚,马履一,等.3种修剪方法对文冠果芽内源激素含量和萌芽成枝的影响[J].西北农林科技大学学报(自然科学版),2017,45(5):101-108.SU M L,WU S,MA L Y,et al.Effects of three pruning methods on endogenous hormone and shoot growth of Xanthoceras sorbifolia Bunge buds[J].Journal of Northwest A&F University (Natural Science Edition),2017,45(5):101-108.
[22] 史绍林.文冠果种子催芽和组织培养研究[D].黑龙江齐齐哈尔:齐齐哈尔大学,2012.
[23] 郝一男,王喜明,丁立军.文冠果油生物柴油副产物粗甘油的精制[J].中国油脂,2011,36(11):80-82.HAO Y N,WANG X M,DING L J.Refining of crude glycerol produced from the production of Xanthoceras sorbiflia Bunge seed oil biodiesel[J].China Oils and Fats,2011,36(11):80-82.
[24] 孙俊.文冠果油的提取及其生物柴油制备工艺研究[D].西安:陕西师范大学,2008.
[25] 白鑫磊,金华,邹吉祥,等.文冠果表型性状的相关性及主成分分析[J].经济林研究,2019,37(1):68-73.BAI X L,JIN H,ZOU J X,et al.Correlation analysis and principal component analysis on phenotypic characteristics in Xanthoceras sorbifolia[J].Nonwood Forest Research,2019,37(1):68-73.
[26] 马新,姜继元,李铭.24个种源文冠果在新疆的生长结实性状分析[J].经济林研究,2019,37(1):61-67.MA X,JIANG J Y,LI M.Analysis on growth and fruiting characteristics of Xanthoceras sorbifolia from 24 provenances in Xinjiang[J].Nonwood Forest Research,2019,37(1):61-67.
[27] 路蒙蒙,韩硕,杨琦,等.文冠果LEC1基因的克隆及表达分析[J].北京林业大学学报,2018,40(1):8-16.LU M M,HAN S,YANG Q,et al.Cloning and expression analysis of LEC1 gene from Xanthoceras sorbifolia embryos[J].Journal of Beijing Forestry University,2018,40(1):8-16.
[28] 赵娜,张媛,李秋琦,等.文冠果FAD2的序列与功能分析[J].北京林业大学学报,2015,37(2):87-93.ZHAO N,ZHANG Y,LI Q Q,et al.Sequence and functional analysis of FAD2 gene from Xanthoceras sorbifolia seeds[J].Journal of Beijing Forestry University,2015,37(2):87-93.
[29] 刘祾悦,阮成江,王莉,等.文冠果种仁油脂脂肪酸形成的多基因协同调控[J].分子植物育种,2019,17(6):1 834-1 842.LIU L Y,RUAN C J,WANG L,et al.Coordinated regulation of multigenes formed by fatty acids in kernel oil of Xanthoceras sorbifolium[J].Molecular Plant Breeding,2019,17(6):1 834-1 842.
[30] 赵翠格.文冠果种子发育过程中油脂累积规律研究[D].北京:北京林业大学,2010.
[31] 张宏平,张晋元,刘玉香,等.桃果实组织结构和超微结构细胞学研究[J].北方园艺,2018,(1):29-34.ZHANG H P,ZHANG J Y,LIU Y X,et al.Histological and ultrastructural cytology of peach fruit[J].Northern Horticulture,2018,(1):29-34.
[32] RUAN Y L,PATRICK J W.The cellular pathway of postphloem sugar transport in developing tomato fruit[J].Planta,1995,196(3):434-444.