不同处理对刺葡萄愈伤组织花青素和原花青素生物合成的影响
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摘要
以刺葡萄2个同质不同型的愈伤组织细胞系为材料,研究不同处理因子对其花青素和原花青素合成能力的影响。试验结果表明,刺葡萄DLR细胞系具有高产花青素和原花青素的能力,在培养至45 d时,其花青素与原花青素的含量均达到最高值,分别为113.85μg/g(FW)和3 562.95μg/g(FW);低温、高温、肉桂酸、壳聚糖、紫外线、黑暗和KT等因子,对DLR细胞系花青素和原花青素的累积表现出不同的效应;其中,4℃低温处理下,DLR细胞花青素的积累效果最佳,比对照提高了2.14倍;壳聚糖处理下,原花青素积累效果最佳,是对照的2.84倍。刺葡萄DLW细胞系不具有或极低的花青素和原花青素合成能力,处理因子调控对其作用效果不明显。本研究的结论为进一步进行刺葡萄细胞花青素和原花青素合成调控奠定了基础。
Using two homogenous genotype spine grape cell lines with different phenotype as materials, the effects of different treatment factors on the biosynthesis of anthocyanins and proanthocyanidins were investigated in this experiment. The results showed that DLR cell line had the potential of high-yield anthocyanins and proanthocyanidins. The highest content of anthocyanin and proanthocyanidins, 113.85 μg/g(FW) and 3 562.95 μg/g(FW), respectively, was observed cultured 45 days after. The accumulation effects of anthocyanin and proanthocyanidins in the DLR cells were various under different treatment factors, such as low temperature, high temperature, cinnamic acid, chitosan, ultraviolet, dark, KT(kinetin) and etc. The accumulation effect of anthocyanin in DLR cells was optimal cultured at 4 ℃, which was 2.14 times higher than that of the control. The accumulation effect of proanthocyanidins in DLR cells was optimal under chitosan treatment, which was 2.84 times higher than that of the control. The DLW cell line couldn't synthesize anthocyanins or proanthocyanidins, and the regulation effect of treatment factors was not obvious. The conclusion of this study laid a foundation for the regulation of anthocyanin and proanthocyanidins biosynthesis in spine grape cells.
Using two homogenous genotype spine grape cell lines with different phenotype as materials, the effects of different treatment factors on the biosynthesis of anthocyanins and proanthocyanidins were investigated in this experiment. The results showed that DLR cell line had the potential of high-yield anthocyanins and proanthocyanidins. The highest content of anthocyanin and proanthocyanidins, 113.85 μg/g(FW) and 3 562.95 μg/g(FW), respectively, was observed cultured 45 days after. The accumulation effects of anthocyanin and proanthocyanidins in the DLR cells were various under different treatment factors, such as low temperature, high temperature, cinnamic acid, chitosan, ultraviolet, dark, KT(kinetin) and etc. The accumulation effect of anthocyanin in DLR cells was optimal cultured at 4 ℃, which was 2.14 times higher than that of the control. The accumulation effect of proanthocyanidins in DLR cells was optimal under chitosan treatment, which was 2.84 times higher than that of the control. The DLW cell line couldn't synthesize anthocyanins or proanthocyanidins, and the regulation effect of treatment factors was not obvious. The conclusion of this study laid a foundation for the regulation of anthocyanin and proanthocyanidins biosynthesis in spine grape cells.
引文
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[21]刘冉,王振宇,李婷婷,等.壳聚糖对红松幼苗多酚积累和抗氧化防御酶的诱导作用[J].植物生态学报,2014,38(7):749-756.
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[23]景艳军,林荣呈.我国植物光信号转导研究进展概述[J].植物学报,2017,52(3):257-270.
[24]Jeong S T,Goto-Yamamoto N,Kobayashi S,et al.Effects of plant hormones and shading on the accumulation of anthocyanins and the expression of anthocyanin biosynthetic genes in grape berry skins[J].Plant Science,2004,167(2):247-252.
[25]Su N,Wu Q,Qi N,et al.Effect of partial shading treatments on anthocyanin synthesis in the hypocotyls of soybean sprouts under UV-A irradiation[J].Plant Growth Regulation,2017,36(1):50-59.
[26]Zhu H,Li X,Zhai W,et al.Effects of low light on photosynthetic properties,antioxidant enzyme activity,and anthocyanin accumulation in purple pak-choi(Brassica campestris ssp.Chinensis Makino)[J].PLo S One,2017,12(6):e0179305.
[27]Zoratti L,Karppinen K,Escobar A L,et al.Light-controlled flavonoid biosynthesis in fruits[J].Frontiers Plant Science,2014,5:534.
[28]Zhao Y,Dong W,Wang K,et al.Differential sensitivity of fruit pigmentation to ultraviolet light between two peach cultivars[J].Frontiers Plant Science,2017,8:1 552.
[29]赵文军,张迪,马丽娟,等.原花青素的生物合成途径、功能基因和代谢工程[J].植物生理学通讯,2009,45(5):509-519.
[30]宫硖,薛静,张晓东.植物花青素合成途径中的调控基因研究进展[J].生物技术进展,2011,1(6):381-390.
[2]王晨,房经贵,曹雪,等.葡萄中原花青素的代谢[J].中国农学通报,2009,25(9):169-173.
[3]Yousuf B,Gul K,Wani A A,et al.Health benefits of anthocyanins and their encapsulation for potential use in food systems:a review[J].Critical Reviews in Food Science and Nutrition,2016,56(13):2 223-2 230.
[4]Li S,Xu M,Niu Q,et al.Efficacy of procyanidins against in vivo cellular oxidative damage:a systematic review and meta-analysis[J].PLoS One,2015,10(10):e0139455.
[5]Eghbaliferiz S,Iranshahi M.Prooxidant activity of polyphenols,flavonoids,anthocyanins and carotenoids:updated review of mechanisms and catalyzing metals[J].Phytotherapy Research,2016,30(9):1 379-1 391.
[6]徐孟川,李述刚,丁玉松,等.葡萄籽原花青素对砷诱导人肝细胞HL-7702损伤的保护作用及其机制研究[J].食品科学,2018,39(3):176-181.
[7]Wei H,Chen X,Zong X,et al.Comparative transcriptome analysis of genes involved in anthocyanin biosynthesis in the red and yellow fruits of sweet Cherry(Prunusavium L.)[J].PLoS One,2015,10(3):e0121164.
[8]Liu M,Song C,Chi M,et al.The effects of light and ethylene and their interaction on the regulation of proanthocyanidin and anthocyanin synthesis in the skins of Vitisvinifera berries[J].Plant Growth Regulation,2016,79(3):377-390.
[9]Isah T,Umar S,Mujib A,et al.Secondary metabolism of pharmaceuticals in the plant in vitro cultures:strategies,approaches,and limitations to achieving higher yield[J].Plant Cell,Tissue Organ Culture,2018,132:239.
[10]曲均革,虞星炬,张卫,等.前体饲喂、诱导子和光照联合使用对葡萄细胞培养合成花青素的影响[J].生物工程学报,2006,22(2):299-305.
[11]杜晓映.葡萄细胞的悬浮培养及诱导子对悬浮细胞多酚产量的研究[D].杨凌:西北农林科技大学,2008:39-44.
[12]赖呈纯,范丽华,黄贤贵,等.刺葡萄幼胚愈伤组织诱导及其高产原花青素细胞系筛选[J].植物生理学报,2014,50(11):1 683-1 691.
[13]Holton T A,Cornish E C.Genetics and biochemistry of anthocyanin biosynthesis[J].Plant Cell,1995,7(7):1 071-1 083.
[14]贾赵东,马佩勇,边小峰,等.植物花青素合成代谢途径及其分子调控[J].西北植物学报,2014,34(7):1 496-1 506.
[15]Wang N,Zhang Z,Jiang S,et al.Synergistic effects of light and temperature on anthocyanin biosynthesis in callus cultures of red-fleshed apple(Malus sieversii f.niedzwetzkyana)[J].Plant Cell,Tissue and Organ Culture,2016,127(1):217-227.
[16]Lo Piero A R,Puglisi I,Rapisarda P,et al.Anthocyanins accumulation and related gene expression in red orange fruit induced by low temperature storage[J].Journal of Agricultural Food and Chemistry,2005,53:9 083-9 088.
[17]Nozaki K,Takamura T,Fukai S.Effects of high temperature on flower colour and anthocyanin content in pink flower genotypes of greenhouse chrysanthemum(Chrysanthemum morifolium Ramat.)[J].Journal of Horticultural Science and Biotechnology,2006,(81):728-734.
[18]黄鸿曼,袁利兵,彭志红,等.花青素的生物合成与环境调控研究进展[J].湖南农业科学,2011(13):118-120.
[19]Mori K,Goto-Yamamoto N,Kitayama M,et al.Loss of anthocyanins in red-wine grape under high temperature[J].Journal of Experimental Botany,2007,58(68):1 935-1 945.
[20]董妍玲,潘学武,赵小虎,等.壳聚糖诱导子促进喜树培养细胞喜树碱积累的研究[J].安徽农业科学,2010,38(29):16 151-16 153.
[21]刘冉,王振宇,李婷婷,等.壳聚糖对红松幼苗多酚积累和抗氧化防御酶的诱导作用[J].植物生态学报,2014,38(7):749-756.
[22]季梅,尹德明,楼红祥.壳聚糖对地钱细胞生长和酚类次生代谢的影响[J].山东农业大学学报(医学版),2007,45(10):1 025-1 029.
[23]景艳军,林荣呈.我国植物光信号转导研究进展概述[J].植物学报,2017,52(3):257-270.
[24]Jeong S T,Goto-Yamamoto N,Kobayashi S,et al.Effects of plant hormones and shading on the accumulation of anthocyanins and the expression of anthocyanin biosynthetic genes in grape berry skins[J].Plant Science,2004,167(2):247-252.
[25]Su N,Wu Q,Qi N,et al.Effect of partial shading treatments on anthocyanin synthesis in the hypocotyls of soybean sprouts under UV-A irradiation[J].Plant Growth Regulation,2017,36(1):50-59.
[26]Zhu H,Li X,Zhai W,et al.Effects of low light on photosynthetic properties,antioxidant enzyme activity,and anthocyanin accumulation in purple pak-choi(Brassica campestris ssp.Chinensis Makino)[J].PLo S One,2017,12(6):e0179305.
[27]Zoratti L,Karppinen K,Escobar A L,et al.Light-controlled flavonoid biosynthesis in fruits[J].Frontiers Plant Science,2014,5:534.
[28]Zhao Y,Dong W,Wang K,et al.Differential sensitivity of fruit pigmentation to ultraviolet light between two peach cultivars[J].Frontiers Plant Science,2017,8:1 552.
[29]赵文军,张迪,马丽娟,等.原花青素的生物合成途径、功能基因和代谢工程[J].植物生理学通讯,2009,45(5):509-519.
[30]宫硖,薛静,张晓东.植物花青素合成途径中的调控基因研究进展[J].生物技术进展,2011,1(6):381-390.
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