花青素代谢对陆地棉叶片和纤维色泽呈现的影响

doi:10.3864/j.issn.0578-1752.2021.09.003

摘要/Abstract

摘要：

【目的】棉花作为一种重要的经济作物和油料作物,其叶片和纤维均可积累色素物质,呈现不同颜色。叶绿素、类胡萝卜素和花青素的含量及其比例是棉花叶片呈色的主要原因,而棕色纤维中主要色素成分为花青素单体氧化聚合而成的原花青素及其衍生物。通过分析陆地棉不同的叶色突变体叶片和纤维中的花青素含量,花青素和原花青素合成途径中关键基因的表达,探究棉花叶片和纤维颜色呈现与花青素合成的关系,为叶色突变体的利用和彩色棉纤维色泽的改良奠定基础。【方法】通过测定21个陆地棉叶色突变体的叶片花青素含量,根据叶色突变体叶片、纤维颜色和花青素含量差异,筛选了其中6个典型的棉花叶色突变体作为研究材料,比较叶片和纤维（开花后15 d）中的花青素含量,分析花青素含量与叶片、纤维颜色呈现的关系;同时检测叶片及不同发育时期纤维（开花后5、10、15和20 d）中花青素合成关键基因GhCHS和原花青素合成途径关键基因GhLAR和GhANR的表达水平,分析目标基因对叶片和纤维颜色呈现的影响。【结果】21个陆地棉叶色突变体叶片中的花青素含量差异显著,呈现紫红色或紫色的叶片花青素含量高。在筛选的6个陆地棉叶色突变体及其对照叶片和不同发育时期纤维中,叶片花青素含量显著高于纤维,棕色纤维的花青素含量显著高于白色纤维。叶片中,GhCHS表达量较高,而GhANR和GhLAR表达量较低,花青素积累与颜色呈现与其表达量没有显著的相关性;而在纤维中,GhANR和GhLAR在棕色纤维的表达量极显著高于白色纤维中,且主要集中在纤维发育的5—15 DPA高表达。【结论】陆地棉叶片和纤维的颜色呈现均与花青素含量有关,紫色及紫红色叶片以及棕色纤维中花青素含量高,但纤维颜色的形成与棉花叶片颜色呈现没有显著的相关性,其花青素含量与原花青素合成途径的关键基因GhANR和GhLAR表达水平直接相关,表明棉花叶片和纤维中的呈色机制不一致,原花青素主要在纤维中积累显色。

关键词: 陆地棉, 叶色突变体, 纤维色泽, 花青素, 基因表达

Abstract:

【Objective】Cotton is an important economic and oil crop. Both its leaves and fibers can accumulate pigments and present different colors. Studies have confirmed that chlorophyll, carotenoids, and anthocyanin are the main pigments in cotton leaves and their relative proportion changes leaf colors. While proanthocyanidins and their derivatives, which are oligomeric and polymeric products from anthocyanidins, are thought to be responsible for the color formation of brown fibers. This article intends to explore the relationship between the color of leaves and fibers in upland cotton through analyzing the anthocyanidin content and gene expression level in the anthocyanin biosynthesis pathway in different leaf color mutants. The result will help to lay the foundation for the utilization of leaf color mutants and the improvement of the color of colored cotton fibers. 【Method】In this experiment, the anthocyanidin contents in leaves of 21 upland cotton leaf color mutants were detected. According to the leaf and fiber color as well as the anthocyanidin content level, 6 cotton leaf color mutants were selected as materials to measure the anthocyanidin level in leaves and fibers at 15 days post anthesis to analyze the relationship between anthocyanidin content and the leaf or fiber color. Then the expression levels of GhCHS, GhLAR and GhANR in leaves and fibers at different developmental stages (5, 10, 15, 20 DPA) were measured to analyze the influence of target genes on the color formation of leaves and fibers. 【Result】The anthocyanidin content in the leaves of 21 leaf color mutants of G. hirsutum was significantly different, and the purple or fuchsia leaves had higher anthocyanidin content. In the selected six leaf color mutants, the anthocyanidin content in leaves was significantly higher than that in fibers, and brown fibers accumulated more anthocyanidins than white fibers. Compared to GhCHS, GhANR and GhLAR expressed at a very low level in leaves, and no significant correlation was found between leaf color and their expression level. While in fibers, the expressions of GhANR and GhLAR were obviously higher in brown fibers than in white fibers, especially in fibers of 5 DPA to 15 DPA. 【Conclusion】Anthocyanins played important roles in color formation of both leaves and fibers in upland cotton. Purple-red or purple leaves and brown fibers accumulated more anthocyanidins, while the formation of fiber color did not directly correlate with leaf color. In fibers, the contents of anthocyanidins directly related to the expression levels of GhANR and GhLAR, indicating that coloration mechanism of cotton leaves and fibers was not exactly the same, and proanthocyanidins mainly accumulated in fibers.

Key words: Gossypium hirsutum L., leaf color mutant, fiber color, anthocyanin, gene expression

袁景丽,郑红丽,梁先利,梅俊,余东亮,孙玉强,柯丽萍. 花青素代谢对陆地棉叶片和纤维色泽呈现的影响[J]. 中国农业科学, 2021, 54(9): 1846-1855.

YUAN JingLi,ZHENG HongLi,LIANG XianLi,MEI Jun,YU DongLiang,SUN YuQiang,KE LiPing. Influence of Anthocyanin Biosynthesis on Leaf and Fiber Color of Gossypium hirsutum L.[J]. Scientia Agricultura Sinica, 2021, 54(9): 1846-1855.

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参考文献 36

[1]	TIAN J, SHEN H, ZHANG J, SONG T, YAO Y. Characteristics of chalcone synthase promoters from different leaf-color malus crabapple cultivars. Scientia Horticulturae, 2011,129:449-458.
[2]	LIN Q H, ZHONG Q Z, ZHANG Z H. Comparative transcriptome analysis of genes involved in anthocyanin biosynthesis in the pink-white and red fruits of Chinese bayberry (Morella rubra). Scientia Horticulturae, 2019,250:278-286.
[3]	BRADSHAW H D, SCHEMSKE D W. Allele substitution at a flower colour locus produces a pollinator shift in monkeyflowers. Nature, 2003,426(6963):176-178.
[4]	SILVA V O, FREITAS A A, MA A, QUINA F H. Chemistry and photochemistry of natural plant pigments: The anthocyanins. Journal of Physical Organic Chemistry, 2016,29:594-599.
[5]	BUENO J M, SAEZ-PLAZA P, RAMOS-ESCUDERO F, MUÑOZ A M, NAVAS M J, ASUERO G A. Analysis and antioxidant capacity of anthocyanin pigments. Part I: General considerations concerning polyphenols and flavonoids. Critical Reviews in Analytical Chemistry, 2012,42:126-151.
[6]	HELDT H W, PIECHULLA B. Phenylpropanoids comprise a multitude of plant secondary metabolites and cell wall components// Plant Biochemistry. 4th ed. ELSEVIER: Academic Press, 2011: 431-449.
[7]	彭祖茂, 邓梦雅, 严虞虞, 朱丽, 张协光. 植物中花青素含量测定及种类分布研究. 食品研究与开发, 2018,39(17):100-104.
	PENG Z M, DENG M Y, YAN Y Y, ZHU L, ZHANG X G. Study on the determination of anthocyanin content and its species distribution in plants. Journal of food research and development, 2018,39(17):100-104. (in Chinese)
[8]	AHMED N U, PARK J I, JUNG H J, HUR Y, NOU I S. Anthocyanin biosynthesis for cold and freezing stress tolerance and desirable color in Brassica rapa. Functional & Integrative Genomics, 2015,15:383-394.
[9]	AHMED N U, PARK J I, JUNG H J, YANG T J, HUR Y, NOU I S. Characterization of dihydro flavonol 4-reductase (DFR) genes and their association with cold and freezing stress in Brassica rapa. Gene, 2014,550(1):46-55.
[10]	AZUMA A, YAKUSHIJI H, KOSHITA Y, KOBAYASHI S. Flavonoid biosynthesis-related genes in grape skin are differentially regulated by temperature and light conditions. Planta, 2012,236:1067-1080.
[11]	LI Z, WANG J, ZHANG X, XU L. Comparative transcriptome analysis of Anthurium “Albama” and its anthocyanin-loss mutant. PLoS ONE, 2015,10(3):e0119027-1-e0119027-20.
[12]	HONG Y, TANG X, HUANG H, ZHANG Y, DAI S. Transcriptomic analyses reveal species-specific light-induced anthocyanin biosynthesis in chrysanthemum. BMC Genomics, 2015,16:202.
[13]	韩科厅, 赵莉, 唐杏姣, 胡可, 戴思兰. 菊花花青素苷合成关键基因表达与花色表型的关系. 园艺学报, 2012,39(3):516-524.
	HAN K T, ZHAO L, TANG X J, HU K, DAI S L. The relationship between the expression of key genes in anthocyanin biosynthesis and the color of chrysanthemum. Acta Horticulturae Sinica, 2012,39(3):516-524. (in Chinese)
[14]	BAN Y, HONDA C, HATSUYAMA Y, IGARASHI M, BESSHO H, MORIGUCHI T. Isolation and functional analysis of a MYB transcription factor gene that is a key regulator for the development of red coloration in apple skin. Plant Cell Physiology, 2007,48(7):958-970.
[15]	BURBULIS I E, WINKEL-SHIRLEY B. Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Poceedings of the National Academy of Sciences of the United States of America, 1999,96(22):12929-12934.
[16]	KOES R E, QUATTROCCHIO F, MOL J N. The flavonoid biosynthetic pathway in plants: Function and evolution. Bioessays, 1994,16(2):123-132.
[17]	DEVIC M, GUILLEMINOT J, DEBEAUJON I, BECHTOLD N, BENSAUDE E, KOORNNEEF M, PELLETIER G, DELSENY M. The BANYULS gene encodes a DFR‐like protein and is a marker of early seed coat development. The Plant Journal, 1999,19(4):387-398.
[18]	FURUKAWA T, ESHIMA A, KOUYA M, TAKIO S, TAKANO H, ONO K. Coordinate expression of genes involved in catechin biosynthesis in Polygonum hydropiper cells. Plant Cell Reports, 2002,21(4):385-389.
[19]	XIE D Y, SHARMA S B, DIXON R A. Anthocyanidin reductases from Medicago truncatula and Arabidopsis thaliana. Archives of Biochemistry and Biophysics, 2004,422(1):100-102.
[20]	HAN Y, VIMOLMANGKANG S, SORIA-GUERRA R E, KORBAN S. Introduction of apple ANR genes into tobacco inhibits expression of both CHI and DFR genes in flowers, leading to loss of anthocyanin. Journal of Experimental Botany, 2012,63(7):2437-2447.
[21]	LIU H F, LUO C, SONG W, SHEN H T, LI G L, HE Z G, CHEN W G, CAO Y Y, HUANG F, TANG S W, HONG P, ZHAO E F, ZHU J B, HE D J, WANG S M, HUO G Y, LIU H L. Flavonoid biosynthesis controls fiber color in naturally colored cotton. PeerJ, 2018,6(10):e4537.
[22]	XIAO Y H, ZHANG Z S, YIN M H, LUO M, LI X B, HOU L, PEI Y. Cotton flavonoid structural genes related to the pigmentation in brown fibers. Biochemical and Biophysical Research Communications, 2007,358(1):73-78.
[23]	郭帅, 郭倩瑜, 郭红彦. 五种绿叶和彩叶树种光合色素含量的动态变化. 安徽农学通报, 2011,17:38-44.
	GUO S, GUO Q Y, GUO H Y. Dynamic changes of photosynthetic pigment contents in five green and colored leaf trees. Anhui Agricultural Science Bulletin, 2011,17:38-44. (In Chinese)
[24]	FENG H, LI Y, WANG S, ZHANG L, LIU Y, XUE F, SUN Y, WANG Y, SUN J. Molecular analysis of proanthocyanins related to pigmentation in brown cotton fiber (Gossypium hirsutum L.). Journal of Experimental Botany, 2014,65(20):5759-5769.
[25]	YATSU L Y, ESPELIE K E, KOLATTUKUDY P E. Ultrastructural and chemical evidence that the cell wall of green cotton fiber is suberized. Plant Physiology, 1983,73(2):521-524.
[26]	SCHMUTZ A, JENNY T, RYSER U. A caffeoyl-fatty acid-glycerol ester from wax associated with green cotton fiber suberin. Phytochemistry, 1994,36(6):1343-1346.
[27]	SCHMUTZ A, BUCHALA A J, RYSER U. Changing the dimensions of suberin lamellae of green cotton fibers with a specific inhibitor of the endoplasmic reticulum-associated fatty acid elongates. Plant Physiology, 1996,110(2):403-411.
[28]	FENG H, YANG Y, SUN S, LI Y, ZHANG L, TIAN J, ZHU Q, FENG Z, ZHU H, SUN J. Molecular analysis of caffeoyl residues related to pigmentation in green cotton fibers. Journal of Experimental Botany, 2017,68(16):4559-4569.
[29]	GIUSTI M M, WROLSTAD R E. Characterization and measurement of anthocyanins by UV-Visible spectroscopy//GIUSTI M M, WROLSTAD R E, eds., Current protocols in food analytical chemistry, John Wiley and Sons, Inc., Hoboken, 2001, F1.2.1.-F1.2.13.
[30]	GAO J F, SHEN L, YUAN J L, ZHENG H L, SU Q S, YANG W G, ZHANG L Q, NNAEMEKA E V, SUN J, KE L P, SUN Y Q. Functional analysis of GhCHS, GhANR and GhLAR in colored fiber formation of Gossypium hirsutum L. BMC Plant Biology, 2019,19(1):455.
[31]	MA D, SUN D, WANG C, LI Y, GUO T. Expression of flavonoid biosynthesis genes andaccumulation of flavonoid in wheat leaves in response to drought stress. Plant Physiology and Biochemistry, 2014,80:60-66.
[32]	ZHAO C L, CHEN Z J, BAI X S, DING C, LONG T J, WEI F G, MIAO K R. Structure-activity relationships of anthocyanidin glycosylation. Molecular Diversity, 2014,18(3):687-700.
[33]	KONG J M, CHIA L S, GOH N K, CHIA T F, BROUILLARD R. Analysis and biological activities of anthocyanins. Phytochemistry, 2003,64(5):923-933.
[34]	吴华玲, 何玉媚, 李家贤, 陈栋, 黄华林, 乔小燕, 刘军. 11个红紫芽茶树新品系的芽叶特性和生化成分研究. 植物遗传资源学报, 2012,13(1):42-47.
	WU H L, HE Y M, LI J X, CHEN D, HUANG H L, QIAO X Y, LIU J. Shoot traits and biological compositions among eleven new tea germplasms with reddish violet shoots. Journal of Plant Genetic Resources, 2012,13(1):42-47. (in Chinese)
[35]	SAURE M C. External control of anthocyanin formation in apple. Scientia Horticulturae, 1990,42(3):181-218.
[36]	FENG H, TIAN X, LIU Y, LI Y, ZHANG X, JONES B J, SUN Y, SUN J. Analysis of flavonoids and the flavonoid structural genes in brown fiber of upland cotton. PLoS ONE, 2013,8(3):e58820.

样本编号 ID	名称 Name	原产地 Place of origin	叶色 Leaf color	纤维颜色 Fiber color
1	红叶白絮 Hongyebaixu	中国山西省 Shanxi Province, China	紫 Purple	白 White
2	石河子822 Shihezi 822	中国新疆建设兵团Xinjiang Construction Corps, China	绿/紫Green/purple	白 White
3	送兴红叶B Songxinghongye B	中国河北Hebei, China	红Red	白 White
4	红叶棕絮 Hongyezongxu	中国山西Shanxi, China	绿Green	棕 Brown
5	锦9-70 Jin 9-70	中国辽宁Liaoning, China	绿Green	白 White
6	紫花棉 Zihuamian	中国辽宁Liaoning, China	绿/紫Green/purple	棕 Brown
7	红叶鸡脚 Hongyejijiao	中国四川Sichuan, China	红Red	白 White
8	安徽红桃棉 Hongtao	中国安徽Anhui, China	红Red	白 White
9	LA877	美国 U.S.A	绿/红Green/purple	白 White
10	抗红叶 Kanghongye	中国四川Sichuan, China	红Red	白 White
11	小红叶 Xiaohongye	中国河南Henan, China	红Red	白 White
12	红叶花苞棉 Hongyehuabaomian	中国河南Henan, China	红Red	白 White
13	矮红株 Aihongzhu	中国江苏Jiangsu, China	紫Purple	白 White
14	芽黄棉1号 Yahuangmian1	中国湖北Hubei, China	黄/绿Yellow/green	白 White
15	花斑叶Huabanye	中国河南Henan, China	斑驳Mottled leaf	白 White
16	斑叶棉 Banyemian	中国河南Henan, China	斑驳Mottled leaf	白 White
17	红鸡脚柳苞 Hongjijiaoliubao	中国贵州 Guizhou, China	紫Purple	白 White
18	红槿矮 Hongjinai	中国湖北 Hubei, China	绿Green	白 White
19	观赏棉120 Guanshangmian120	中国河北Hebei, China	中红Middle red	白 White
20	贵池红叶 Guichihongye	中国安徽Anhui, China	紫Purple	白 White
21	HS2	中国浙江Zhejiang, China	紫Purple	白 White

引物名称 Primer name	序列 Primer sequence (5′-3′)	用途 Use
T-GhCHS-F	GCTGTTACCTTTCGTGGACC	基因表达分析 Gene expression analysis
T-GhCHS-R	TCGCACCATCACTATCTGGC	基因表达分析 Gene expression analysis
T-GhANR-F	GACGTTGGCTGAAAAGGCAG	基因表达分析 Gene expression analysis
T-GhANR-R	CAGAAAAACATGGGCTCGGC	基因表达分析 Gene expression analysis
T-GhLAR-F	GCATCTTGGACCAGCTCAGT	基因表达分析 Gene expression analysis
T-GhLAR-R	GTGTTGTCATGGTAGGGCCA	基因表达分析 Gene expression analysis
T-GhUBQ7-F	GAAGGCATTCCACCTGACCAAC	内参基因 Reference gene
T-GhUBQ7-R	CTTGACCTTCTTCTTCTTGTGCTTG	内参基因 Reference gene