,1, 李付广,2Effects of Different Red and Blue Ratios on the Somatic Embryogenesis and Plant Regeneration of Cotton
WEI Xi1,2, WANG QianHua2, GE XiaoYang2, CHEN YanLi2, DING YanPeng2, ZHAO MingZhe,1, LI FuGuang,2通讯作者:
责任编辑: 李莉
收稿日期:2018-11-29接受日期:2019-01-27网络出版日期:2019-03-16
| 基金资助: |
Received:2018-11-29Accepted:2019-01-27Online:2019-03-16
作者简介 About authors
魏喜,E-mail： chinaweixi521@163.com。
王倩华,E-mail： qianhuahehe@163.com。 魏喜和王倩华为同等贡献作者。
摘要
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魏喜, 王倩华, 葛晓阳, 陈艳丽, 丁颜朋, 赵明哲, 李付广. 不同红蓝配比的光质对棉花体细胞胚胎发生 和植株再生的影响[J]. 中国农业科学, 2019, 52(6): 968-980 doi:10.3864/j.issn.0578-1752.2019.06.002
WEI Xi, WANG QianHua, GE XiaoYang, CHEN YanLi, DING YanPeng, ZHAO MingZhe, LI FuGuang.
0 引言
【研究意义】棉花是全球最重要的油料作物和经济作物之一,其产量和纤维品质关系着国计民生[1]。世界上超过80多个国家种植棉花,中国是世界上最大的原棉生产国和消费国。但由于受到非生物胁迫、病虫害等不利因素的制约,严重影响棉花的产量和纤维品质[2]。因此,通过棉花基因工程导入相关基因,增强棉花抗逆性和抗病虫害,从而提高棉花产量和品质是必需的[3]。将转基因技术应用到棉花育种,弥补了传统育种方式周期长等不足[4]。然而体细胞胚胎发生是一个非常复杂的动态变化的过程,涉及到各种生物大分子的合成及运输、信号传导和能量变化等[5]。这种复杂性影响了体细胞胚胎发生因素的多样性,其中光照是影响体细胞胚胎发生的重要因素之一,对其生长和分化有重要影响[6]。体细胞胚胎发生基本上都是在日光灯下培养,然而随着科技的进步,不同发光颜色的LED灯应用到生产实践中,探究不同光质组合对体胚发生的影响,为提高棉花体胚发生能力和加快转基因进程奠定基础。然而针对环境可控棉花组织培养系统,红蓝光作为人工光源对体细胞胚胎发生的研究鲜有报道。【前人研究进展】研究表明,光合色素吸收红光和蓝光的能力比其他波长的能力强,红光影响茎的伸长、根冠比、叶绿素含量和光合作用[7,8],然而蓝光影响向光性、下胚轴伸长、叶片膨胀、气孔开放、酶合成、叶绿体运动和基因表达等[9,10]。虽然蓝光和红光有各自不同的功能,但是单独使用红光或蓝光不能满足植物生长发育的要求[11]。研究表明,与单色光相比,红蓝混合光能提高光合速率和茎的质量[12,13]。红蓝光作为人工光源的研究大多集中于不同光强和光质对植物生长和品质的影响：闻婧等[14]研究表明,红蓝配比为B﹕R=1﹕8的LED光源明显提高莴苣光合速率和品质以及降低耗电量;王君等[15]发现红蓝配比为B﹕R=1﹕1,光强为300 μmol·m-2·s-1处理组合提高生菜的光能、电能利用效率及干重累积量;合理光质组合和光照强度可促进烤烟幼苗根系与叶片的生长[16]。棉花体细胞胚胎发生和植株再生的过程包括愈伤诱导、胚性愈伤诱导、体细胞胚诱导和植株再生,其中,胚性愈伤分化阶段是非常重要的阶段,胚性愈伤分化的时间和状态,以及环境因素都可以影响这一过程。目前,棉花体细胞胚胎发生的研究主要集中在寻求合适的棉花组织培养体系,以及挖掘调控棉花体细胞胚胎发生能力的关键基因。例如,董合忠等[17]研究发现2,4-D和KT同时添加对愈伤诱导的效果明显高于单独添加激素的效果;于娅等[18]研究发现,激素由高到低的继代可明显提高胚性愈伤分化率,IAA/KT比例为1﹕1—1﹕6,胚性愈伤分化率最高为50.22%。ZHENG等[3]研究认为高分化率材料生长素运输基因PIN7和信号转导通路基因SHY2表达量比难分化或不分化的材料表达量要高。YANG等[19]研究发现棉花体细胞胚胎发生过程中生长素合成、运输、代谢、信号应答等相关基因表达均有不同程度的变化,进一步证实了棉花体细胞胚胎发生这一复杂性。XIAO等[20]在拟南芥中过表达棉花GhWUSs,结果发现过表达GhWUSs明显促进拟南芥芽再生能力,并且LEC1、FUS3、ABI3和CLV3的表达量明显上升。GE等[21]利用iTRAQ技术分离鉴定与棉花体细胞胚发育相关的关键通路,发现压力响应、激素合成和信号转导、糖和能量代谢、蛋白代谢、细胞壁代谢、细胞转运和脂类物质转运等通路在调控棉花体细胞胚发育方面发挥着重要的作用。【本研究切入点】目前,有关棉花体细胞胚胎发生的研究大多集中在寻找合理培养基配方和关键功能基因的挖掘,不同光质配比对棉花体细胞胚胎发生过程影响的研究很少,特别是不同红蓝配比的光质对棉花体细胞胚胎发生的影响更少。【拟解决的关键问题】本研究分别设置不同红蓝配比为B﹕R=1﹕1、B﹕R=3﹕1、B﹕R= 1﹕3和DL的光质组合,探索其对棉花体细胞胚胎发生各个阶段的影响,为提高棉花体细胞胚胎发生能力和构建大量快速的获得棉花再生植株的工厂化生产实践体系奠定基础。1 材料与方法
1.1 植物材料和生长条件
选用的棉花外植体材料是CCRI12,由中国农业科学院棉花研究所提供,试验于2016—2018年依托中棉所转基因课题组棉花规模化转基因技术平台开展。光源为光强和光质可精确调控的红蓝LED人工光源。将难分化材料CCRI12种子播种于中棉所试验田,收获时挑选成熟度良好的棉花种子,将CCRI12种子消过毒后放在无菌苗培养基上[3],其中,基本培养基为MS和B5,这里简称MSB培养基[22],为了方便使用,首先将其配成母液MSB培养基（大量元素MSⅠ：50 mL·L-1、微量元素MSⅡ：5 mL·L-1、B5维生素：5 mL·L-1和铁盐：5 mL·L-1）,固体培养基每升再加5.5 g琼脂粉,121℃,灭菌14 min。在MSB基础培养基上配置一系列的培养基：无菌苗培养基（大量元素MSⅠ 50 mL、蔗糖28 g、琼脂粉5.5 g,用超纯水充分溶解后定容到1 L,pH自然）。棉花体细胞胚胎发生一般要经历4个过程：愈伤组织诱导培养基、胚性愈伤分化培养基、体细胞胚诱导培养基和再生苗培养基,分别简称为CIM、ECDM、EIM和RIM,4种培养基的主要成分如表1。
Table 1
表1
表1棉花体细胞胚胎发生阶段各培养基成分
Table 1
| 名称 Name | 基础培养基 MSB (mL·L-1) | 蔗糖 Sucrose (g·L-1) | 葡萄糖 Glucose (g·L-1) | 6-糠氨基嘌呤 Kinetin (KT) (mg·L-1) | 2,4-二氯苯氧乙酸 2,4-D (mg·L-1) | 吲哚乙酸 IAA (mg·L-1) | 6-苄氨基腺嘌呤 6-BA (mg·L-1) | 琼脂 Gelose (g·L-1) |
|---|---|---|---|---|---|---|---|---|
| 愈伤诱导培养基CIM | 65 | 28 | 0.1 | 0.1 | 2.2 | |||
| 胚性愈伤诱导培养基ECDM | 65 | 28 | 0.15 | 0.05 | 2.2 | |||
| 体细胞胚诱导培养基EIM | 65 | 28 | 0.1 | 0.2 | 2.2 | |||
| 再生苗诱导培养基RIM | 65 | 25 | 0.05 | 0.05 |
新窗口打开|下载CSV
适宜的培养条件是成功获得再生棉花苗的必备条件。将配制好的培养基分装到100 mL三角瓶中,每瓶50 mL,高温高压灭菌后,将无菌苗CCRI12下胚轴切成数个长度为5—7 mm的大小。所有培养物均置于（28±2）℃。
1.2 试验设计
设置的不同红蓝配比光质分别是B﹕R=1﹕1、B﹕R=3﹕1、B﹕R=1﹕3和DL 4种组合,其中B﹕R= 1﹕1、B﹕R=3﹕1和B﹕R=1﹕3是不同的红蓝配比,DL光质处理是白光处理。红蓝光的波峰分别为660和460 nm。DL光质处理的光强设置在30—50 μmol·m-2s-1。采用不同红蓝配比的LED光源,其设置依据是不同蓝红配比光源对植物的干重和鲜重[23],以及叶片的光合速率[24]产生不同的影响。每个处理分别设置3个重复。1.3 仪器和测量方法
采用频谱分析仪（HR-350,台湾,高雄）测量光谱分布,LED光源板（XM-LEDX12T8D,中国,江苏,南京）上的红蓝光光强大小是通过调节供电电流和电灯泡的数量来确定的。CPR = (W2-W1)/W1,其中,W1为外植体培养前的初始重量;W2为外植体培养一段时间后的重量;CPR（callus proliferation rate）：指的是愈伤增殖率,即单位时间内外植体重量变化量除以外植体初始重量。
愈伤增殖量是指培养一段时间后愈伤增加的重量;胚性愈伤分化率是指由愈伤组织经过再分化成胚性愈伤组织的愈伤数量与再分化前愈伤块总数量的比值;体细胞胚数量是指胚性愈伤组织发育成不同状态的体细胞胚的总数,包括球形胚、心形胚、鱼雷胚和子叶胚等;生根率是指发育有根的下胚轴数量与下胚轴总数的比值。
叶绿素含量的测定：称取样品0.1—0.2 g（鲜重,FW）,用8 mL 80%的丙酮作为提取液,4℃,黑暗条件下放置24 h,充分混匀,直到叶片褪色至白色为之。将混合物充分离心后采用分光光度计UV-2550（Shimadzu, Kyoto, Japan）测定646.6、663.6和450 nm的吸光度,根据吸光度的值计算叶绿素的浓度。叶绿素浓度的计算方法与前人研究一致[25]。
1.4 数据处理
采用Origin 8.5、SSPS20.0等软件进行数据处理和分析。采用最小显著差数法（the least significant difference,LSD）进行显著性检验（P<0.05）。2 结果
2.1 棉花体细胞胚胎发生过程
在棉花组织培养过程中,棉花下胚轴被认为是作为棉花体细胞胚胎发生的理想外植体。不同光质处理棉花体细胞胚胎发生一般要经历4个阶段（图1-A）。分别为愈伤诱导阶段诱导出愈伤（callus formation）、胚性愈伤诱导阶段诱导出胚性愈伤组织（embryogenic callus,ECs）、体细胞胚阶段诱导出体细胞胚（somatic embryos maturation,SEs）和再生棉花苗阶段。7 d苗龄大小的CCRI12棉花下胚轴作为外植体,将下胚轴用无菌刀片切成5—7 mm（图1-B）,混合均匀后放在愈伤诱导培养基上诱导愈伤的形成,随着诱导天数的增加,愈伤的状态由早期愈伤（图1-C）逐渐转变为后期愈伤状态（图1-D）,后期愈伤状态蓬松,含水量丰富。将愈伤转移到胚性愈伤组织分化培养基诱导胚性愈伤（ECs）的形成。培养一段时间后,会诱导出大量的ECs（图1-E）。接着将胚性愈伤转移到体细胞胚诱导培养基,诱导体细胞胚（SEs）的形成,根据体细胞胚发育状态的不同,将体细胞胚分为球形胚（图1-F）、心形胚（图1-G）、鱼雷胚（图1-H）和子叶胚（图1-I）。最后将这些体细胞胚再转移到再生植株诱导培养基上,继续培养,直到获得再生棉花植株（图1-J）。图1
新窗口打开|下载原图ZIP|生成PPT图1棉花体细胞胚胎发生过程
A：棉花体细胞胚胎发生的流程图;B：5—7 mm大小的棉花下胚轴切段;C：早期愈伤;D：后期愈伤;E：胚性愈伤组织;F：球形胚;G：心形胚;H：鱼雷胚;I：子叶胚;J：再生棉花植株
Fig. 1Cotton somatic embryogenesis
A: Flow chart of cotton somatic embryogenesis; B: 5-7 mm cotton hypocotyl segments; C: Early callus; D: Late callus; E: Embryogenic callus; F: Globular embryo; G: Heart-shaped embryo; H: Torpedo embryo; I: Cotyledon embryo; J: Regenerated cotton plant
2.2 不同光质组合影响愈伤的增殖
不同的光质组合对棉花愈伤组织的增殖有不同的影响。CPR这个参数被用来衡量愈伤的增殖情况。CCRI12下胚轴在CIM上培养30 d后（图2-A）,B﹕R=1﹕1、B﹕R=3﹕1、B﹕R=1﹕3和DL 4种光组合诱导出的愈伤在颜色和质地上没有差异,但是愈伤块的大小有明显的差异,B﹕R=1﹕3处理后的愈伤块是最小的,B﹕R=1﹕1和B﹕R=3﹕1处理后的愈伤块比较大。根据这种愈伤状态,分别统计4种光处理的愈伤增殖率（图2-B）,发现B﹕R=1﹕1处理的CPR最大,显著高于B﹕R=3﹕1处理,极显著高于B﹕R=1﹕3和DL处理。B﹕R=3﹕1处理的CPR显著高于B﹕R=1﹕3和DL处理,B﹕R=1﹕3和DL光处理之间的CPR不显著。综上说明B﹕R=1﹕1和 B﹕R=3﹕1光质组合促进愈伤的增殖,B﹕R=1﹕3处理对愈伤增殖贡献较小;不同光质组合在愈伤后期对愈伤的颜色和质地有显著影响。图2
新窗口打开|下载原图ZIP|生成PPT图2不同光质处理对CPR的影响
A：4种不同红蓝配比下愈伤组织的表型;B：4种不同光质处理下对愈伤增殖率的统计,采用最小显著差数法（LSD）进行显著性检验（P<0.05）;不同小写字母表示LSD检验各组间两两比较的统计学差异（P<0.05）。下同
Fig. 2Effect of different light quality treatments on CPR
A: Callus phenotype under four different red and blue ratios; B: Callus proliferation rate under four different light quality treatments was carried out by the least significant difference (LSD) method (P < 0.05). Different lower-case letters in B denote statistical differences of pairwise comparisons among each group of bars according to LSD test (P<0.05). The same as below
2.3 不同光质组合对胚性愈伤阶段的影响
2.3.1 不同光质组合在胚性愈伤诱导阶段对未分化愈伤的影响 愈伤诱导阶段主要是诱导外植体的体细胞脱分化形成大量能再分化为胚性愈伤的细胞团。图3-A为不同光质组合在CIM上培养30 d后的愈伤转移到ECDM上继续培养45 d后的表型。B﹕R=1﹕1、B﹕R=3﹕1、B﹕R=1﹕3和DL 4种光组合中,B﹕R=1﹕1、B﹕R=3﹕1和DL处理后未分化愈伤块的体积都明显大于B﹕R=1﹕3处理后的未分化愈伤块的体积。为了明确不同光质组合对未分化愈伤的影响,在ECDM上培养45 d后,统计未分化愈伤的增殖量（图3-B）。B﹕R=1﹕1、B﹕R=3﹕1和DL处理的光质组合的愈伤增殖量分别是7.85、6.51和4.08 g,但是B﹕R=1﹕3处理的光质组合的愈伤增殖量只有1.06 g。与B﹕R=1﹕1和B﹕R= 3﹕1处理相比,B﹕R=1﹕3光质组合处理的未分化愈伤增殖量差异显著,但是与DL处理光质组合的愈伤增殖量相比差异不显著。因此,B﹕R=1﹕1和B﹕R=3﹕1处理在胚性愈伤诱导阶段促进未分化愈伤的大量增殖,而B﹕R=1﹕3光质组合处理对未分化愈伤增殖的贡献较小。图3
新窗口打开|下载原图ZIP|生成PPT图3不同光质处理对未分化愈伤的影响
A：4种不同红蓝配比下未分化愈伤组织的表型;B：4种不同光质处理下对未分化愈伤增殖量的统计
Fig. 3Effects of different light quality treatments on undifferentiated callus
A: Phenotype of undifferentiated callus under four different red-blue ratios; B: Statistical analysis of the proliferation of undifferentiated callus under four different light quality treatments
2.3.2 不同光质组合对棉花胚性愈伤组织诱导的影响 在不同光质组合诱导胚性愈伤阶段,除了上述增殖不分化的愈伤外,另一部分愈伤分化为胚性愈伤。图4-A是在胚性愈伤诱导阶段培养45 d时的状态,其中B﹕R=1﹕1和B﹕R=3﹕1处理的愈伤表现出大量增殖的表型,只有部分愈伤分化为胚性愈伤（ECs）,胚性愈伤分化率分别是0.25和0.16。B﹕R=1﹕3处理和DL处理的光质组合能诱导大量胚性愈伤（图4-A红色椭圆）,胚性愈伤分化率分别是0.70和0.56。B﹕R=1﹕3处理的光质组合的胚性愈伤分化率极显著高于B﹕R=1﹕1和B﹕R=3﹕1处理的光质组合,显著高于DL处理。综上所述,与白光处理相比,B﹕R=1﹕3处理的光质组合显著促进愈伤分化成胚性愈伤,而B﹕R=1﹕1和B﹕R=3﹕1处理的光质组合抑制胚性愈伤组织的形成。
图4
新窗口打开|下载原图ZIP|生成PPT图4不同光质处理对棉花胚性愈伤组织诱导的影响
A：4种不同红蓝配比下胚性愈伤组织的表型;B：4种不同光质处理下对胚性愈伤分化率的统计。ECs：胚性愈伤
Fig. 4Effects of different light quality treatments on embryogenic callus induction in cotton
A: Phenotypes of hypocotylic callus with four different red and blue ratios; B: Statistical analysis of differentiation rate of embryonic callus with four different light quality treatments. ECs: Embryogenic callus
2.4 不同光质组合对棉花体细胞胚诱导阶段的影响
2.4.1 不同光质组合对棉花体细胞胚数量的影响 B﹕R=1﹕1、B﹕R=3﹕1、B﹕R=1﹕3和DL光质组合都能诱导出胚性愈伤,将黄绿色、颗粒状的胚性愈伤组织放置在体细胞胚诱导培养基上进一步诱导,胚性愈伤的颜色逐渐变绿,首先发育成原胚然后依次发育成球形胚、心形胚、鱼雷胚和子叶胚。每个处理各选50瓶,每隔一个月继代一次,直到体细胞胚的出现。图5-A为不同光质组合下诱导出的体细胞胚,统计球形胚,、心形胚、鱼雷胚和子叶胚总数量（图5-B）。3次重复的试验结果表明,B﹕R=1﹕3处理组合的体细胞胚总数显著高于B﹕R=1﹕1和B﹕R=3﹕1处理组合诱导体细胞胚的数量。虽然B﹕R=1﹕3处理组合的体细胞胚总数与DL处理的体细胞胚总数相比差异不显著,但是B﹕R=1﹕3处理组合的体细胞胚总数也比DL处理的体细胞胚总数多出50个。综上所述,B﹕R=1﹕3处理组合对体细胞胚诱导具有促进作用,B﹕R=1﹕1和B﹕R=3﹕1处理对体细胞胚诱导有抑制作用。图5
新窗口打开|下载原图ZIP|生成PPT图5不同光质处理对棉花体细胞胚诱导阶段的影响
A：4种不同红蓝配比下体细胞胚的表型;B：4种不同光质处理下对体细胞胚数量的统计
Fig. 5Effects of different light quality treatments on the induction stage of cotton embryoids
A: The phenotype of somatic embryos under four different red and blue ratios; B: The statistics of somatic embryos under four different light quality treatments
2.4.2 不同光质组合对棉花体细胞胚形成下胚轴的影响 在体细胞胚诱导培养基上继代培养1—2次后,将体细胞胚转移到再生植株诱导培养基上培养直到长出棉花再生苗。分别在不同光质组合处理B﹕R=1﹕1、B﹕R=3﹕1、B﹕R=1﹕3和DL下继续培养,当培养15 d后体细胞胚就诱导出下胚轴（图6-A）。图6-B中可以看出B﹕R=1﹕3和DL的光质组合下诱导出下胚轴的数量明显多于B﹕R=1﹕1和B﹕R=3﹕1光质组合诱导出的下胚轴数量。B﹕R=1﹕3和DL光质组合处理之间的下胚轴数量没有显著差异,同样的B﹕R=1﹕1和B﹕R=3﹕1光质组合处理之间下胚轴的数量没有显著差异。总之,B﹕R=1﹕1和B﹕R=3﹕1处理不利于体细胞胚形成下胚轴,而B﹕R=1﹕3和DL的光质组合处理促进体细胞胚形成下胚轴。
图6
新窗口打开|下载原图ZIP|生成PPT图6不同光质处理对棉花体细胞胚长下胚轴的影响
A：4种不同红蓝配比下体细胞胚诱导出下胚轴的表型;B：4种不同光质处理下对体细胞胚诱导出下胚轴的统计
Fig. 6Effects of different light quality treatments on the number of hypocotyls from SEs
2.5 不同光质组合对棉花再生苗诱导阶段的影响
2.5.1 不同光质组合对下胚轴生根的影响 将下胚轴在再生植株诱导培养基上继代1次后诱导不定根的发生（图7-A）,B﹕R=1﹕1、B﹕R=3﹕1、B﹕R=1﹕3和DL都能诱导出大量不定根。且4种不同光质组合对下胚轴长根的影响有明显差异（图7-B）,B﹕R=1﹕1处理的下胚轴生根率明显高于B﹕R=3﹕1、B﹕R=1﹕3和DL不同光质组合处理,且差异显著;而B﹕R=3﹕1、B﹕R=1﹕3和DL处理之间的下胚轴生根率差异不显著。图7
新窗口打开|下载原图ZIP|生成PPT图7不同光质处理对棉花下胚轴生根的影响
A：4种不同红蓝配比下生根情况的表型;B：4种不同光质处理下对生根率的统计
Fig. 7Effects of different light quality treatments on rooting of cotton hypocotyls
A: Phenotypes of rooting under four different ratios of red and blue; B: Statistics of rooting rates under four different light quality treatments
2.5.2 不同光质组合对棉花再生苗和植株高度的影响 图8-A是不同光质组合处理下棉花再生苗的表型,在B﹕R=1﹕1和B﹕R=1﹕3不同光质组合处理的再生棉花苗植株高大,B﹕R=3﹕1和DL不同光质组合处理的棉花植株矮小。经统计成苗数量（图8-B）,B﹕R=1﹕3光质组合下成苗数量明显高于B﹕R=3﹕1和DL处理下成苗数,且差异显著;B﹕R=1﹕3光质组合成苗数与B﹕R=1﹕1光质组合下的成苗数差异不显著。B﹕R=1﹕1光质组合诱导的成苗数量显著高于B﹕R=3﹕1处理后诱导的成苗数量,但是与DL光质处理诱导的成苗数量相比差异不显著。B﹕R=3﹕1光质组合处理与DL光质处理相比,对再生苗的诱导数量差异不显著。统计不同光质配比下棉花植株高度（图8-C）,B﹕R=1﹕1和B﹕R=1﹕3光质组合下棉花植株的高度分别是14.58和13.93 cm,显著高于B﹕R=3﹕1与DL,但二者之间差异不显著。B﹕R=3﹕1与DL光质配比下棉花植株高度分别是5.53和5.95 cm,差异不显著。综上,B﹕R=1﹕1和B﹕R=1﹕3的光质组合促进再生苗的产生和发育。
图8
新窗口打开|下载原图ZIP|生成PPT图8不同光质组合对棉花再生苗和植株高度的影响
A：4种不同红蓝配比下再生棉花植株的表型;B：4种不同光质处理下对下胚轴成苗数的统计;C：4种不同光质处理下对下胚轴成苗数的统计
Fig. 8Effects of different light qualities on cotton seedling regeneration and plant height
A: The phenotype of regenerated cotton plants under four different red-blue ratios; B: the statistics of seedling number of hypocotyl under four different light quality treatments; C: the statistics of seedling number of hypocotyl under four different light quality treatments
2.5.3 不同光质组合对棉花再生苗叶绿素浓度的影响 图9-A是在不同光质组合处理下,棉花再生苗的叶片的表型,B﹕R=1﹕1和B﹕R=1﹕3不同光质处理下,棉花叶片深绿,但是B﹕R=1﹕1和B﹕R=3﹕1不同光质处理下,棉花叶片嫩绿;整体上B﹕R=1﹕3不同光质处理下棉花叶片体积大,颜色深。通过对诱导的棉花再生苗进行叶绿素含量的测定（图9-B）。B﹕R=1﹕3的光质组合下叶绿素浓度最高,与B﹕R=1﹕1光质组合相比,差异不显著,但是与B﹕R=3﹕1的光质组合处理相比,差异显著;与DL光质组合相比,差异显著。综上,与DL光质处理相比,B﹕R=1﹕1、B﹕R=3﹕1和B﹕R=1﹕3处理组合都有利于叶绿素的合成,但是B﹕R=1﹕3处理组合促进叶绿素的合成效果最明显。
图9
新窗口打开|下载原图ZIP|生成PPT图9不同光质组合对棉花再生苗叶绿素浓度的影响
A：4种不同红蓝配比下再生棉花植株叶片的表型;B：4种不同光质处理下叶绿素浓度的统计
Fig. 9Effects of different light qualities on chlorophyll concentration of cotton regenerated seedlings
A: Leaf phenotype of regenerated cotton under four different red-blue ratios; B: Statistical analysis of chlorophyll concentration under four different light quality treatments
3 讨论
体细胞胚胎的发生方式为直接发生和间接发生2种途径。一些报道[26]认为陆地棉体细胞胚胎发生大多是通过间接发生途径完成,然而也有研究表明陆地棉的植株再生有可能通过体细胞胚胎直接发生,但是成功的概率非常[27,28]。下胚轴是作为体细胞胚胎发生的理想外植体,体细胞胚发生中愈伤起始的组织位置可能不完全一致,既有近表层愈伤组织内部的报道,也有内外同时起源的报道,可能与培养方式有关[29]。3.1 不同红蓝配比的光质对愈伤组织形成和分化的影响
愈伤组织是由棉花下胚轴外植体经过脱分化形成的一类细胞团。B﹕R=1﹕1和B﹕R=3﹕1红蓝光配比下愈伤表现出大量增殖的状态,但愈伤分化成胚性愈伤严重受阻;而B﹕R=1﹕3下愈伤增殖率最低,显著抑制愈伤的增殖,但胚性愈伤的分化率显著提高。B﹕R=1﹕1和B﹕R=3﹕1光质组合的愈伤大量增殖而难分化,B﹕R=1﹕3光质组合表现出愈伤增殖较小,容易分化成胚性愈伤,说明愈伤适度增殖有利于分化成胚性愈伤,这与XU[30]的研究一致。棉花通过体细胞胚胎发生获得再生棉花是一个非常困难的过程,周期长而且分化率非常低,基因型限制严重。大部分棉花品种都只能形成愈伤,不能分化成胚性愈伤,因此胚性愈伤的诱导在棉花体细胞胚胎发生过程中起着重要的作用。B﹕R=1﹕1和B﹕R=3﹕1光质组合表现出大量增殖而难分化,B﹕R=1﹕3和DL的光质组合愈伤增殖率小而分化率比较高,说明不同光质组合对愈伤细胞多能性获得及分化成胚性愈伤的能力的影响不同,这与 ARACELI等[31]结论一致。从体细胞脱分化成愈伤组织,再分化成胚性愈伤组织的过程,使这些细胞转变成具有发育成体细胞胚潜力的细胞团,其间涉及到细胞生理变化、新陈代谢和基因的表达等[32]。NHUT等[33]研究表明B﹕R=2﹕3的光质组合对油茶体细胞胚发生有利,促进油茶植物的再生;而D"ONOFRIO等[6]研究表明柑橘在红光条件下体细胞胚胎发生能力是最高的,在红蓝混合光和正常日光灯条件下体细胞胚胎发生能力是降低的。以上结果表明,不同植物对光的响应是不同的,导致不同光质组合对体细胞胚胎发生的调控能力有很大差异。3.2 不同红蓝配比的光质对体细胞胚发育的影响
在胚性愈伤发育成体细胞胚的过程中,各种生理和化学因素发挥着重要的作用,包括光质、激素等。B﹕R=1﹕3光质组合能明显促进体细胞数量的增加,而B﹕R=1﹕1和B﹕R=3﹕1光质组合对诱导体细胞的产生有一定的抑制作用,导致体细胞数量的降低。B﹕R=1﹕3的光质组合可能精确调控了细胞内生长素、细胞分裂素、脱落酸和茉莉酸的变化,这些激素的变化诱导体细胞胚胎发生的Marker基因的表达,进而调控体细胞胚的发育。相似的研究指出,LEC1、LEC2、FUS3通过调控赤霉素和脱落酸的动态平衡来诱导和维持体细胞胚的发育[21]。HERINGER等[34]利用甘蔗的芽尖分生组织作为外植体研究不同红蓝光质组合对体细胞胚胎发生的影响,发现低比例的蓝光抑制甘蔗植株的转化,而蓝光和红光以及远红光相结合有利于叶片和根的伸长,增加叶片数和叶绿素的含量,对不同红蓝光质组合下甘蔗体细胞胚胎发生的蛋白组学分析发现,生长素、PIN的运输蛋白和DNA甲基化伴随着甘蔗的体细胞胚胎发生。3.3 不同红蓝配比的光质对下胚轴的影响
B﹕R=1﹕1和B﹕R=3﹕1光质组合下体细胞胚诱导出下胚轴的数量明显低于B﹕R=1﹕3和DL的处理组合;B﹕R=1﹕1和B﹕R=1﹕3处理组合下诱导出的再生棉花的高度显著高于B﹕R=3﹕1和DL组合。再生棉花植株的数量很大程上取决于诱导出下胚轴的数量和质量。研究表明,蓝光激活油菜素内酯（BR）和赤霉素（GA）信号通路,进而诱导HBI1表达促进下胚轴伸长[35]。不同红蓝配比的光照下CRY的差异表达调控了HBI1的差异表达,影响体细胞胚下胚轴的发育。XU[36]研究表明在红光、蓝光、白光、远红光条件下,BBX21的差异表达直接影响拟南芥下胚轴的长度。B﹕R=1﹕1处理后生根率明显高于B﹕R=1﹕3、B﹕R=3﹕1和DL光质组合,可能的原因是B﹕R=1﹕1组合引起侧根发育相关的基因WOX5、WOX7、WOX11和WOX12的过量表达[37]。3.4 不同红蓝配比的光质对叶绿素浓度的影响
B﹕R=1﹕3、B﹕R=3﹕1和B﹕R=1﹕1处理后叶绿素浓度明显高于DL组合。研究表明LED 2种波峰和R/B对叶用莴苣叶绿素含量影响不同[38]。LEDA型（波峰为660 nm的红光LED与450 nm的蓝光LED组成）光源处理的叶绿素a、b和（a+b）含量显著高于LEDB型光源（波峰为630 nm的红光LED与460 nm的蓝光LED组成）处理[14]。YE等[39]研究表明蓝光增加金线莲叶绿素的含量与蓝光和红光以及CK相比。不同植物对不同红蓝配比光质的响应能力不同,组织培养过程中使用的红蓝光的光强度和红蓝光质配比不一致,这些因素可能导致了不同植物在红蓝光处理下的光合作用的能力不同,最终导致叶绿素含量不同。4 结论
在棉花体细胞胚胎发生过程中,不同发育阶段对光的响应能力有差异,愈伤增殖阶段对蓝光的响应最敏感,但过度的愈伤增殖不利于胚性愈伤的产生;胚性愈伤、体细胞胚诱导和苗再生阶段对红光响应最敏感。采用B﹕R=1﹕3的光质组合有利于棉花体细胞胚胎的发生。参考文献 原文顺序
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DOI:10.3969/j.issn.1002-7807.2000.06.010URL [本文引用: 1]
棉纤维是重要的纺织工业原料。随着人民群众对衣着要求的普遍提高 ,纺工部门引入高效快速的纺纱技术 ,对棉花品种的总体纤维品质 ,提出了更高的要求。目前国内外棉花育种家除了利用常规技术外 ,还大规模开展了棉纤维品质改良的分子育种研究。本文综述了国内外利用克隆出的棉纤维发育基因以及外源基因开展棉纤维品质改良的转基因育种和棉纤维优质基因的分子标记筛选和辅助选择育种的研究进展。同时 ,根据我国目前棉花品种品质水平、生产状况 ,提出了棉花品质育种策略。
DOI:10.3969/j.issn.1002-7807.2000.06.010URL [本文引用: 1]
棉纤维是重要的纺织工业原料。随着人民群众对衣着要求的普遍提高 ,纺工部门引入高效快速的纺纱技术 ,对棉花品种的总体纤维品质 ,提出了更高的要求。目前国内外棉花育种家除了利用常规技术外 ,还大规模开展了棉纤维品质改良的分子育种研究。本文综述了国内外利用克隆出的棉纤维发育基因以及外源基因开展棉纤维品质改良的转基因育种和棉纤维优质基因的分子标记筛选和辅助选择育种的研究进展。同时 ,根据我国目前棉花品种品质水平、生产状况 ,提出了棉花品质育种策略。
DOI:10.1023/A:1015673200621URL [本文引用: 1]
Somatic embryogenesis is defined as a process in which a bipolar structure, resembling a zygotic embryo, develops from a non-zygotic cell without vascular connection with the original tissue. Somatic embryos are used for studying regulation of embryo development, but also as a tool for large scale vegetative propagation. Somatic embryogenesis is a multi-step regeneration process starting with formation of proembryogenic masses, followed by somatic embryo formation, maturation, desiccation and plant regeneration. Although great progress has been made in improving the protocols used, it has been revealed that some treatments, coinciding with increased yield of somatic embryos, can cause adverse effects on the embryo quality, thereby impairing germination and ex vitro growth of somatic embryo plants. Accordingly, ex vitro growth of somatic embryo plants is under a cumulative influence of the treatments provided during the in vitro phase. In order to efficiently regulate the formation of plants via somatic embryogenesis it is important to understand how somatic embryos develop and how the development is influenced by different physical and chemical treatments. Such knowledge can be gained through the construction of fate maps representing an adequate number of morphological and molecular markers, specifying critical developmental stages. Based on this fate map, it is possible to make a model of the process. The mechanisms that control cell differentiation during somatic embryogenesis are far from clear. However, secreted, soluble signal molecules play an important role. It has long been observed that conditioned medium from embryogenic cultures can promote embryogenesis. Active components in the conditioned medium include endochitinases, arabinogalactan proteins and lipochitooligosaccharides.
DOI:10.1023/a:1006059615088URL [本文引用: 2]
The effect of light quality on somatic embryogenesis in quince BA 29 was investigated. 2,4-D induced leaves were exposed for 25 days to the following light quality treatments: dark, far-red, far-red+blue, far-red+red, blue, white, red+blue, red. After a further 20 days of white light exposure, somatic embryo production was recorded. Somatic embryogenesis was highest in cultures subjected to red light treatment, and decreased progressively with the transition to red+blue and to white. Overall, embryogenic competence showed a correlation with photoequilibrium. Phytochrome appeared to be inductive although this effect was adversely influenced by the blue absorbing photoreceptor, in particular at low photoequilibrium. Independently of light treatments applied, somatic embryos frequently showed severe morphological abnormalities. Conversion of somatic embryos to plantlets was not observed.
[本文引用: 1]
DOI:10.1007/BF00051588URL [本文引用: 1]
Cultures in vitro of Betula pendula Roth were subjected to light of different spectral qualities. Photosynthetic capacity was highest when the plantlets were exposed to blue light (max recorded photosynthesis, 82 μmol CO 2 dm 612 h 611 ) and lowest when irradiated with light high in red and/or far-red wave lengths (max recorded photosynthesis, 40 μmol CO 2 dm 612 h 611 ). Highest chlorophyll content (2.2 mg dm 612 leaf area) was found in cultures irradiated with blue light, which also enhanced the leaf area. Morphometric analysis of light micrographs showed that the epidermal cell areas were largest in plantlets subjected to blue light and smallest in those subjected to red light. Morphometric analysis of electron micrographs of palisade cells, showed that the functional chloroplast area was largest in chloroplasts of leaves subjected to blue light and smallest in those exposed to red light. We suggest that light quality affects photosynthesis both through effects on the composition of the photosynthetic apparatus and on translocation of carbohydrates from chloroplasts.
DOI:10.1073/pnas.0709189105URL [本文引用: 1]
DOI:10.1016/j.jphotobiol.2009.03.010URLPMID:19410482 [本文引用: 1]
Light quality is thought to affect many plant physiological processes during growth and development, particularly photosynthesis. We examined how light quality influences plant photosynthesis by analyzing changes in photosynthetic parameters and expression levels of some photosynthesis related genes of cucumber ( Cucumis sativus L. cv. Jinyou No. 1) plants. The plants were grown under different light qualities: purple ( P), blue ( B), green ( G), yellow ( Y), red ( R) and white light ( W) of the same photosynthetic photon flux density (PFD) about 350 μmol m 612 s 611 for 5 days. The results show that all plants grown under monochromatic light had reduced growth, CO 2 assimilation rate (Pn) and quantum yield of PSII electron transport (Φ PSII) as compared with plants grown under W, and these reductions were more significant in the plants under G, Y and R. The decrease in Φ PSII is mostly due to the reduction in photochemical quenching (qP). Interestingly, P- and B-grown plants had higher stomatal conductance (Gs), total and initial Rubisco activities and higher transcriptional levels of 10 genes which encode key enzymes in the Calvin cycle together with higher total soluble sugars, sucrose and starch contents as compared with W-grown plants, whereas in G-, Y-, and R-grown plants these parameters declined. Therefore, the reduction in Pn under P and B is likely the result of inactivation of photosystems, whilst under Y, G and R it is caused by, in addition to photosystem inactivation, the closure of stomata and the transcriptional down-regulation of genes for the Calvin cycle enzymes such as rbc L and rca. In conclusion, light quality alters plant photosynthesis by the effects on the activity of photosynthetic apparatus in leaves and the effects on the expression and/or activity of the Calvin cycle enzymes.
DOI:10.1007/s11099-015-0083-8URL [本文引用: 1]
The objective of this study was to evaluate the significance of blue light (B) in the growth and photosynthetic capacity of cucumber. Gas exchange, chlorophyll (Chl) fluorescence kinetics, chloroplast ultrastructure, and leaf growth were investigated to explore the influence of three different light qualities of light emitting diodes (LEDs) on plant morphogenesis and the development of photosynthetic apparatus in cucumber ( Cucumis sativus ) leaves from emergence to full expansion under weak light [50 mol(photon) m 2 s 1 ]. We found that B could significantly increase the leaf area (LA), shoot elongation, Chl a / b , net photosynthetic rate, and stomatal conductance ( g s ). In addition, the comparisons of maximal quantum yield of PSII photochemistry and the photosynthetic performance index between B-, W (white light)-, and R (red light)-grown leaves suggested that B was essential for the development of photosynthetic apparatus under weak light. B-grown leaves had the lowest Chl content under weak light, however, they had well-developed chloroplasts with the highest degree of stacked lamellae and the lowest starch accumulation. This could explain to a considerable extent the highest net photosynthetic rate per Chl unit. The results demonstrated that B optimized photosynthetic performance by improving the photosynthetic rate, increasing LA, and prolonging active photosynthesis duration under low irradiance. Therefore B is necessary to ensure healthy development of chloroplasts and highly efficient photosynthetic functions in cucumbers under a weak light environment. More importantly, our study also provided theoretical and technical support for the development of light environmental control technology.
DOI:10.1016/j.scienta.2012.10.009 [本文引用: 1]
The light sources generally used for in vitro plant cultures are fluorescent lamps. To select a more efficient light source, the present study evaluated the effects of different light qualities on the growth and morphogenesis of rapeseed (Brassica napus L.) cultivar Westar plantlets in vitro. The plantlets were exposed to 60μmolm612s611 photosynthetic photon flux (PPF) for a 12h photoperiod under the following six different light qualities: fluorescent lamps (FL), monochromic blue light-emitting diodes (LEDs) (B), monochromic red LED (R), and three mixtures of B plus R (3:1, 1:1, 1:3) LED. The proliferation rate was greater in plantlets that were cultured under B light than those under FL. The differentiation rate, fresh mass, dry mass, concentration of chlorophyll a, soluble sugar concentration, stem diameter, leaf stomata abaxial surface length, adaxial surface stomata frequency and transplantation survival rate were greater in plantlets that were cultured under B:R=3:1 light than under FL. The concentration of starch and the spongy tissue length were higher in plantlets cultured under R light than those under FL. The B:R=3:1 LED light was suitable for rapeseed plantlet growth in vitro and can be used as a priority light source in the rapeseed culture system according to its differentiation rate, proliferation rate, growth rate, and transplantation survival rate.
DOI:10.1016/j.envexpbot.2015.04.001URL [本文引用: 1]
Light emitting diodes (LEDs) are frequently regarded as a new light source for the production of horticultural crops under closed-type conditions. However, before use of LEDs as the sole source of light can be advanced, plant responses to light quality have to be investigated for important horticultural plants. The objective of the present study was to evaluate cucumber (Cucumis sativus) seedlings physiological responses to different blue (B) and red (R) photon flux (PF) ratios using LEDs. Cucumber seedlings (cv. Cumlaude) were grown in a growth chamber until the second true leaf stage (17 days) with LED lighting and 18-h photoperiod. The treatments consisted of 100 molm 2s 1 photosynthetic photon flux (PPF) with B:R PF ratios of 0B:100R%, 10B:90R%, 30B:70R%, 50B:50R%, 75B:25R%, 100B:0R%. Another treatment consisted of B, green (G) and R PF ratio of 20B:28G:52R%. Peak wavelengths of LEDs were 455nm (B) and 661nm (R) for the in the B:R treatments and 473nm (B), 532nm (G), 660nm (R) in the B:G:R treatment. Hypocotyl length decreased with the increase of B PF up to the 75B:25R% treatment. Hypocotyl length in the 0B:100R% treatment was 164% greater than in the 75B:25R treatment. Plants under the 100B:0R% treatment had unexpected greater plant height, hypocotyl, and epicotyl length than plants under all other treatments. For example, the hypocotyl length under the 100B:0R% was 69% greater than in the 0B:100R treatment and 346% greater than in the 75B:25R% treatment. Leaf area decreased with the increase of B PF when plants were irradiated with the combination of B and R PF. The response of leaf area under the 100B:0R% treatment was unexpected since plants in the 100B:0R% treatment had 48% greater leaf area than plants in the 75B:25R% treatment. Chlorophyll content per leaf area, net photosynthetic rate, and stomatal conductance increased with the increase of B PF. Shoot dry and fresh mass decreased with the increase of B PF when plants were irradiated with the combination of B and R PF. Plants under 0B:100R% had the lowest dry and fresh mass from all the treatments and plants under 100B:0R% showed the greatest fresh mass from all the treatments and equal dry mass as the plants under 10B:90R% treatment. The addition of G PF to the spectrum did not have any influence in cucumber plant responses. For cucumber seedlings, morphological responses influenced plant growth since B PF responses in growth parameters (i.e., dry mass) closely matched those in morphological parameter (i.e., leaf area). More research is needed to find the optimal spectrum for the growth and development of horticultural crops under sole source electrical lighting such as LEDs.
URLMagsci [本文引用: 2]
为了探求适合叶用莴苣生长的节能高效的光源参数，采用红、蓝光波峰组合分别为红光（R）660 nm + 蓝光（B）450 nm的LEDA型和红光（R）630 nm + 蓝光（B）460 nm的LEDB型光源，每种光源设置R/B分别为6、8和10的3种比例，处理叶用莴苣。结果表明：LEDA型与LEDB型光源处理的叶用莴苣光合速率、蒸腾速率、气孔导度、胞间CO2浓度、维生素C、总糖和硝酸盐含量均无显著差异，但均较荧光灯光源（对照）显著提高了光合速率。两种光源不同R/B处理的上述指标变化趋势一致，均表现为R/B = 8时（LEDA2、LEDB2）最优，其中LEDA2处理比荧光灯处理光合速率提高38%，维生素C含量增加8.3%，硝酸盐含量降低9.2%；LEDB2处理时光合速率提高48%，硝酸盐含量降低6.5%。LEDA型光源处理的叶绿素a、b和（a + b）含量显著高于LEDB型光源处理，且分别表现为LEDA3和LEDB2处理的含量最高。此外，对光源装置耗电量的计算表明，LEDB型光源的单株耗电量比LEDA型和荧光灯（对照）分别节省53.3%和27.7%。因此，红、蓝光波峰分别为630 nm + 460 nm的组合LED光源，R/B = 8的条件下，在提高叶用莴苣光合速率和品质以及降低耗电量3个方面体现优势。 <BR>
URLMagsci [本文引用: 2]
为了探求适合叶用莴苣生长的节能高效的光源参数，采用红、蓝光波峰组合分别为红光（R）660 nm + 蓝光（B）450 nm的LEDA型和红光（R）630 nm + 蓝光（B）460 nm的LEDB型光源，每种光源设置R/B分别为6、8和10的3种比例，处理叶用莴苣。结果表明：LEDA型与LEDB型光源处理的叶用莴苣光合速率、蒸腾速率、气孔导度、胞间CO2浓度、维生素C、总糖和硝酸盐含量均无显著差异，但均较荧光灯光源（对照）显著提高了光合速率。两种光源不同R/B处理的上述指标变化趋势一致，均表现为R/B = 8时（LEDA2、LEDB2）最优，其中LEDA2处理比荧光灯处理光合速率提高38%，维生素C含量增加8.3%，硝酸盐含量降低9.2%；LEDB2处理时光合速率提高48%，硝酸盐含量降低6.5%。LEDA型光源处理的叶绿素a、b和（a + b）含量显著高于LEDB型光源处理，且分别表现为LEDA3和LEDB2处理的含量最高。此外，对光源装置耗电量的计算表明，LEDB型光源的单株耗电量比LEDA型和荧光灯（对照）分别节省53.3%和27.7%。因此，红、蓝光波峰分别为630 nm + 460 nm的组合LED光源，R/B = 8的条件下，在提高叶用莴苣光合速率和品质以及降低耗电量3个方面体现优势。 <BR>
DOI:10.11841/j.issn.1007-4333.2016.08.08URL [本文引用: 1]
通过调查红蓝LED光源下生菜的光能利用效率(LUE)和电能利用效率(EUE),以期确定人工光栽培环境下红蓝LED的优化光强参数.本试验LED光源的红蓝光配比设定为1∶1,光强分别为200、300和400μmol/(m2·s)的3个处理(分别以L200、L300和L400表示),生菜定植38 d后收获.结果表明:1)定植至收获期间,对每次间苗所收获生菜的干重以及定植38 d后收获生菜的干重进行累计计算,处理L300下生菜的EUE最大,为0.948％,较处理L200和L400下生菜的EUE分别高5.22％和46.6％;处理L200下生菜的LUE最大,为4.17％,较处理L300和L400下生菜的LUE分别高12.0％和59.3％.2)定植38 d后处理L300下生菜地上部分的鲜干重最大,平均值分别为41.3和2.58 g,较处理L200和L400下地上部分鲜重分别高23.5％和25.2％;地上部分干重分别高28.4％和12.8％.3)处理L300与处理L200相比,定植38d后前者叶片的可溶性糖含量较高,硝酸盐含量较低;但与处理L400之间无显著性差异.综合考虑光能、电能利用效率及干重累积量,推荐300μmol/(m2·s)作为红蓝LED组合光源下生菜生长较优的光照强度.
DOI:10.11841/j.issn.1007-4333.2016.08.08URL [本文引用: 1]
通过调查红蓝LED光源下生菜的光能利用效率(LUE)和电能利用效率(EUE),以期确定人工光栽培环境下红蓝LED的优化光强参数.本试验LED光源的红蓝光配比设定为1∶1,光强分别为200、300和400μmol/(m2·s)的3个处理(分别以L200、L300和L400表示),生菜定植38 d后收获.结果表明:1)定植至收获期间,对每次间苗所收获生菜的干重以及定植38 d后收获生菜的干重进行累计计算,处理L300下生菜的EUE最大,为0.948％,较处理L200和L400下生菜的EUE分别高5.22％和46.6％;处理L200下生菜的LUE最大,为4.17％,较处理L300和L400下生菜的LUE分别高12.0％和59.3％.2)定植38 d后处理L300下生菜地上部分的鲜干重最大,平均值分别为41.3和2.58 g,较处理L200和L400下地上部分鲜重分别高23.5％和25.2％;地上部分干重分别高28.4％和12.8％.3)处理L300与处理L200相比,定植38d后前者叶片的可溶性糖含量较高,硝酸盐含量较低;但与处理L400之间无显著性差异.综合考虑光能、电能利用效率及干重累积量,推荐300μmol/(m2·s)作为红蓝LED组合光源下生菜生长较优的光照强度.
DOI:10.3969/j.issn.1000-6850.2013.22.029URL [本文引用: 1]
In order to explore light supplement technology by floating seeding of flue-cured tobacco in winter in Hunan, it was studied for effect on different proportion of red blue to growth of root and leaf and carbon and nitrogen metabolism and enzyme activity of the tobacco seeding.Tests are separately carried out with the light emitting diodes in proportion of blue:red 1:3 ,blue:red 1:1 and blue:red 3:1,and they are combined into different light intensity of light source of tobacco seeding by fluorescent light.The results showed that light quality and light intensity on the tobacco leaf time had little effect,and the growth of tobacco seedings are best when light intensity reaches above 7200 Lux and the proportion of red:blue 3:1.The proportion of red:blue 3:1 can promote chlorophyll synthesis and growth of tobacco seeding,but the proportion of red:blue 1:3 can affect the growth of root and increase root activity. Blue light intensity of the total light intensity between 5% and 15% in the combined light significantly promoted the growth of the stem of tobacco seeding.With the increasing of light intensity, the processing leaf of tobacco seeding of the proportion of red:blue 3:1 in combination source can significantly increase total soluble sugar content and enzyme activity,and the processing leaf of tobacco seeding of the proportion red:blue 1:3 in combination source can significantly increase protein content and nitrate reductase activity.Light quality and light intensity on chlorothyll synthesis and carbon and nitrogen metabolism of tobacco seeding highly has sensitive regulation.Reasonable compensation can promote the growth of root and leaf of flue-cured tobacco seeding.
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DOI:10.3969/j.issn.1000-6850.2013.22.029URL [本文引用: 1]
In order to explore light supplement technology by floating seeding of flue-cured tobacco in winter in Hunan, it was studied for effect on different proportion of red blue to growth of root and leaf and carbon and nitrogen metabolism and enzyme activity of the tobacco seeding.Tests are separately carried out with the light emitting diodes in proportion of blue:red 1:3 ,blue:red 1:1 and blue:red 3:1,and they are combined into different light intensity of light source of tobacco seeding by fluorescent light.The results showed that light quality and light intensity on the tobacco leaf time had little effect,and the growth of tobacco seedings are best when light intensity reaches above 7200 Lux and the proportion of red:blue 3:1.The proportion of red:blue 3:1 can promote chlorophyll synthesis and growth of tobacco seeding,but the proportion of red:blue 1:3 can affect the growth of root and increase root activity. Blue light intensity of the total light intensity between 5% and 15% in the combined light significantly promoted the growth of the stem of tobacco seeding.With the increasing of light intensity, the processing leaf of tobacco seeding of the proportion of red:blue 3:1 in combination source can significantly increase total soluble sugar content and enzyme activity,and the processing leaf of tobacco seeding of the proportion red:blue 1:3 in combination source can significantly increase protein content and nitrate reductase activity.Light quality and light intensity on chlorothyll synthesis and carbon and nitrogen metabolism of tobacco seeding highly has sensitive regulation.Reasonable compensation can promote the growth of root and leaf of flue-cured tobacco seeding.
DOI:10.3321/j.issn:1000-7091.1993.03.016URLMagsci [本文引用: 1]
诱导棉花下胚轴切段形成愈伤组织需要外源2，4-D，加入激动素（KT）可以增进2，4-D的效果。愈伤组织形成胚性细胞和胚性细胞团的过程需要2，4-D和KT的共同作用。KT可以促进胚性细胞团分化出胚状体，加入2，4-D则会降低KT的效果。采用2，4-D（0.05mg/L）+KT（0.1mg/L）诱导愈伤组织，60天后转移到无激素的液体培养基上振荡培养，得到了大量的胚状体。
DOI:10.3321/j.issn:1000-7091.1993.03.016URLMagsci [本文引用: 1]
诱导棉花下胚轴切段形成愈伤组织需要外源2，4-D，加入激动素（KT）可以增进2，4-D的效果。愈伤组织形成胚性细胞和胚性细胞团的过程需要2，4-D和KT的共同作用。KT可以促进胚性细胞团分化出胚状体，加入2，4-D则会降低KT的效果。采用2，4-D（0.05mg/L）+KT（0.1mg/L）诱导愈伤组织，60天后转移到无激素的液体培养基上振荡培养，得到了大量的胚状体。
DOI:10.3321/j.issn:1000-4025.2004.02.021URL [本文引用: 1]
以陆地棉"中棉所24"为材料进行了全固体体细胞培养,获得了愈伤组织和再生植株。愈伤组织诱导阶段采用0.01IAA+0.01KT+0.012,4-D的培养基效果好,继代时间多为30～50d;激素由高到低的继代可明显提高胚性愈伤分化率,IAA和KT含量均较低,IAA/KT比例为1∶1～1∶6,胚性愈伤最高分化率为50.22%。
DOI:10.3321/j.issn:1000-4025.2004.02.021URL [本文引用: 1]
以陆地棉"中棉所24"为材料进行了全固体体细胞培养,获得了愈伤组织和再生植株。愈伤组织诱导阶段采用0.01IAA+0.01KT+0.012,4-D的培养基效果好,继代时间多为30～50d;激素由高到低的继代可明显提高胚性愈伤分化率,IAA和KT含量均较低,IAA/KT比例为1∶1～1∶6,胚性愈伤最高分化率为50.22%。
DOI:10.1186/1471-2229-12-110URLPMID:22817809 [本文引用: 1]
pAbstract/p pBackground/p pSomatic embryogenesis (SE), by which somatic cells of higher plants can dedifferentiate and reorganize into new plants, is a notable illustration of cell totipotency. However, the precise molecular mechanisms regulating SE remain unclear. To characterize the molecular events of this unique process, transcriptome analysis, in combination with biochemical and histological approaches, were conducted in cotton, a typical plant species in SE. Genome-wide profiling of gene expression allowed the identification of novel molecular markers characteristic of this developmental process./p pResults/p pRNA-Seq was used to identify 5,076 differentially expressed genes during cotton SE. Expression profile and functional assignments of these genes indicated significant transcriptional complexity during this process, associated with morphological, histological changes and endogenous indole-3-acetic acid (IAA) alteration. Bioinformatics analysis showed that the genes were enriched for basic processes such as metabolic pathways and biosynthesis of secondary metabolites. Unigenes were abundant for the functions of protein binding and hydrolase activity. Transcription factor ncoding genes were found to be differentially regulated during SE. The complex pathways of auxin abundance, transport and response with differentially regulated genes revealed that the auxin-related transcripts belonged to IAA biosynthesis, indole-3-butyric acid (IBA) metabolism, IAA conjugate metabolism, auxin transport, auxin-responsive protein/indoleacetic acid-induced protein (Aux/IAA), auxin response factor (ARF), small auxin-up RNA (SAUR), Aux/IAA degradation, and other auxin-related proteins, which allow an intricate system of auxin utilization to achieve multiple purposes in SE. Quantitative real-time PCR (qRT-PCR) was performed on selected genes with different expression patterns and functional assignments were made to demonstrate the utility of RNA-Seq for gene expression profiles during cotton SE./p pConclusion/p pWe report here the first comprehensive analysis of transcriptome dynamics that may serve as a gene expression profile blueprint in cotton SE. Our main goal was to adapt the RNA-Seq technology to this notable development process and to analyse the gene expression profile. Complex auxin signalling pathway and transcription regulation were highlighted. Together with biochemical and histological approaches, this study provides comprehensive gene expression data sets for cotton SE that serve as an important platform resource for further functional studies in plant embryogenesis./p
DOI:10.1016/j.plantsci.2018.02.018URL [本文引用: 1]
The() gene encodes a plant-specific homeodomain-containing transcriptional regulator, which plays important roles during embryogenesis, as well as in the formation of shoot and flower meristems. Here, we isolated two homologues of(),and, from upland cotton (). Domain analysis suggested that the two putative GhWUS proteins contained a highly conserved DNA-binding HOX domain and a WUS-box. Expression profile analysis showed thatwere predominantly expressed during the embryoid stage. Ectopic expression ofincould induce somatic embryo and shoot formation from seedling root tips. Furthermore, in the absence of exogenous hormone, overexpression ofincould promote shoot regeneration from excised roots, and in the presence of exogenous auxin, excised roots expressingcould be induced to produce somatic embryo. In addition, expression of the chimericrepressor in cotton callus inhibited embryogenic callus formation. Our results show thatis an important regulator of somatic embryogenesis and shoot regeneration.
DOI:10.1021/pr500688gURLPMID:25367710 [本文引用: 2]
Somatic embryo development (SED) in upland cotton shows low frequencies of embryo maturation and plantlet regeneration. Progress in increasing the regeneration rate has been limited. Here a global analysis of proteome dynamics between globular and cotyledonary embryos was performed using isobaric tags for relative and absolute quantitation to explore mechanisms underlying SED. Of 6318 proteins identified by a mass spectrometric analysis, 102 proteins were significantly up-regulated and 107 were significantly down-regulated in cotyledonary embryos. The differentially expressed proteins were classified into seven functional categories: stress responses, hormone synthesis and signal transduction, carbohydrate and energy metabolism, protein metabolism, cell wall metabolism, cell transport, and lipid metabolism. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis showed that stress response, hormone homeostasis, and respiration and photosynthesis were involved in SED. Quantitative real-time PCR analysis confirmed the authenticity and accuracy of the proteomic analysis. Treatment of exogenous hormones showed that abscisic acid and jasmonic acid facilitate SED, whereas gibberellic acid inhibits SED and increases abnormal embryo frequency. Thus, global analysis of proteome dynamics reveals that stress response, hormone homeostasis, and respiration and photosynthesis determined cotton SED. The findings of this research improve the understanding of molecular processes, especially environmental stress response, involved in cotton SED.
[D].
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DOI:10.1016/j.scienta.2003.10.003URL [本文引用: 1]
Node cuttings of in vitro chrysanthemum (1.0 cm) were transplanted on MS basal media supplemented with 30 g l 611 sucrose and cultured for 35 days under six different light qualities: fluorescent (FL), blue LEDs (B), red LEDs (R), red and blue LEDs (RB), red and far-red LEDs (RFr), and blue and far-red LEDs (BFr). Net photosynthetic rate was highest under RB followed by FL and lowest under BFr and B. Fresh weight, dry weight and leaf area were also greatest under FL and RB, while decreased the most under BFr. Greatest stem length was obtained under R and RFr but the stem elongation was due to internode elongation, particularly, the third internode where elongation was almost double the first internode elongation. No difference was observed in the number of nodes among light treatments. Light quality also affected the size and number of leaf stomata. Plantlets under RB, where the growth was greatest (except stem elongation), had the smallest number (56.4 mm 612) and largest leaf stomata ( 32.9 μm×44.7 μm), whereas those under BFr had the least growth, largest number (98.7 mm 612) and smallest leaf stomata ( 24.3 μm×31.7 μm). In conclusion, R and RFr resulted in the highest stem elongation but also resulted in stem fragility due to excessive elongation of the third internode. Shoot growth excluding stem elongation was greatest under RB and FL. To increase stem length without stem fragility in chrysanthemum, it may be required to adjust the ratio of red light when mixed with blue or florescent.
DOI:10.1007/s11240-006-9191-2URL [本文引用: 1]
The aim of the study was to establish whether the quantity and the quality of light affect growth and development of Withania somnifera plantlets. We have studied growth and histo-physiological parameters [stomatal characteristics, chloroplastic pigments concentrations, photosynthesis, and transpiration (E)] of W. somnifera plantlets regenerated under various light intensities, or monochromatic light or under a mixture of two colors of light in tissue culture conditions. Plantlets grown under a photon flux density (PFD) of 3002μmol02m -2 02s -1 showed greater growth and development than those raised under other PFDs. Chlorophylls and carotenoids, numbers of stomata, rate of photosynthesis (P N ) and transpiration (E), stomatal conductance (g s ), and water use efficiency (WUE) increased with increasing the PFD up to 6002μmol02m -2 02s -1 . Light quality also affected plantlets growth and physiology. Highest growth was observed under fluorescent and in a mixture of blue and red light. Very few stomata were developed in any of the monochromatic light but under fluorescent or under a mixture of two colors stomatal numbers increased. Similarly, g s , E, P N, and WUE were also higher under fluorescent light and under a mixture of red and blue light. Regressional analysis showed a linear relationship between P N ( r 2 02=0270) and g s and between E ( r 2 02=020.95) and g s . In conclusion, both the quality and the quantity of light affect growth of plantlets, development of stomata and physiological responses differently depending on the intensity and the wavelength of light.
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本文围绕陆地棉组织细胞培养中的组织培养、单细胞培养、原生质体培养、染色体变异和体细胞抗性筛选等方面的若干问题,在介绍国内外研究进展的同时,着重结合我们的研究工作进行了探讨。
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URL [本文引用: 1]
本文围绕陆地棉组织细胞培养中的组织培养、单细胞培养、原生质体培养、染色体变异和体细胞抗性筛选等方面的若干问题,在介绍国内外研究进展的同时,着重结合我们的研究工作进行了探讨。
DOI:10.1007/s002990000281URL [本文引用: 1]
Abstract  The development of a regeneration system from cotton stomatal guard cells directly on epidermal strips is described. The most important factors affecting embryogenic callus initiation in both of the varieties tested (Coker 312 and 315) were the source of the epidermal tissue, including plant age (4–5 months old), the developmental stage of the flower (opening flower stage) from which bracts were obtained, the composition of the culture medium and light irradiance. The flower developmental stage was critical for callus formation, which was observed only from bracts obtained from opening flowers. In addition, epidermal strips excised from the bract basal region were more responsive in culture than those obtained from the top region. Improved callus initiation was obtained on epidermal strips which had their cuticle in contact with the culture medium. Light irradiance was a limiting factor for embryogenic callus formation, which was observed only in calluses cultured under the lower light irradiance (15.8 μmol m–2 s–1). Somatic embryogenesis was observed on callus cultures subcultured consecutively to a culture medium containing naphthalene acetic acid (10.7 μM) and isopentenyladenine (4.9 μM). Histodifferentiation of somatic embryos was improved on a medium containing naphthaleneacetic acid (8.1 μM)+isopentenyladenine (2.5 μM) and abscisic acid (0.19–0.38 μM). Somatic embryo germination and plantlet development were obtained using established protocols with few modifications. On average, one fully developed plant was obtained from the culture of circa 100 epidermal strips in both cultivars. Keywords Epidermis - Embryogenesis - Regeneration
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本文在回顾棉花细胞和组织培养历史的基础上,评述棉花体细胞胚发生的细胞学和组织学以及植株再生的条件和途径方面的研究进展。
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URL [本文引用: 1]
本文在回顾棉花细胞和组织培养历史的基础上,评述棉花体细胞胚发生的细胞学和组织学以及植株再生的条件和途径方面的研究进展。
DOI:10.3321/j.issn:1000-4025.2009.03.033URL [本文引用: 1]
棉花是一种较难通过体细胞胚发生完成植株再生的作物,而体细胞胚发生是限制棉花基因工程和细胞工程得以广泛应用的主要因素.研究者从不同角度探讨了棉花体细胞胚发生机理并取得了很大的进展,为棉花的遗传转化提供了重要的技术支持,同时成功建立了一些棉花植株再生体系.本文分别从基因型限制、胚胎学、生理生化变化、分子机理等方面阐述了近年来棉花体细胞发生机制的研究进展,并讨论了本研究领域目前存在的主要问题及相应对策.
DOI:10.3321/j.issn:1000-4025.2009.03.033URL [本文引用: 1]
棉花是一种较难通过体细胞胚发生完成植株再生的作物,而体细胞胚发生是限制棉花基因工程和细胞工程得以广泛应用的主要因素.研究者从不同角度探讨了棉花体细胞胚发生机理并取得了很大的进展,为棉花的遗传转化提供了重要的技术支持,同时成功建立了一些棉花植株再生体系.本文分别从基因型限制、胚胎学、生理生化变化、分子机理等方面阐述了近年来棉花体细胞发生机制的研究进展,并讨论了本研究领域目前存在的主要问题及相应对策.
DOI:10.1111/pbi.12947URLPMID:29754405 [本文引用: 1]
DOI:10.1007/s11240-010-9815-4URL [本文引用: 1]
Somatic embryogenesis in Agave tequilana Weber var. Azul was affected by the interaction between the light regimes applied during the induction phase and the expression phase. When embryogenic calli was exposed to white or red light during the expression phase, an average of two germinated embryos per explant was obtained regardless of the light treatment used for callus induction. Conversely, the highest number of germinated embryos, an average of 18 per explant, was obtained when applying either white or red light during the induction phase and then wide-spectrum light during the expression phase. Culture medium had also a great influence in this process, with embryo germination being reduced by up to 70%, depending on the light treatment, when using Schenk and Hildebrandt (SH) medium instead of Murashige and Skoog (MS) medium.
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DOI:10.1080/13102818.2014.1000210URLPMID:09 [本文引用: 1]
In recent years, LED (light-emitting diode) has been the subject of research within the field of plant growth and development. However, there has been little discussion about using LED in vitro cultures of Panax vietnamensis, one of the important medicinal plants belonging to the Panax genus. This study examines the influence of various LED lamps on callus growth and plant formation of P. vietnamensis. Results show significant differences in growth and development, as various light conditions were suitable for different stages. Callus of 70 mg in fresh weight cultured under yellow LEDs resulted in growth of 1197 mg in fresh weight and 91.7 mg of dry weight, within a period of three months. The most effective plant formation was obtained when embryogenic calli were cultured under the combination of 60% red LED and 40% blue LED with an average of 11.21 plantlets per explant; the shoot clump fresh weight and dry weight were of 1147 and 127 mg, respectively, and the average plant height was 3.1 cm. It was also shown that this light condition was the most efficient for P. vietnamensis in vitro plant growth and development. This study provided additional evidence regarding the influence of different LEDs on ginsenoside production applying high-performance liquid chromatography (HPLC) analysis with photo-diode array (PDA) detection at ultraviolet (UV) wavelength 203 nm. The highest MR2 content was recorded when plants maintained under 20% red LED combined with 80% blue LED. However, the highest Rg1 and Rb1 content was found under fluorescent light. The results presented might provide new strategies using LEDs for adequate micropropagation protocols of P. vietnamensis.
DOI:10.1007/s11738-017-2349-1URL [本文引用: 1]
Sugarcane is a renewable energy source that provides an alternative to fossil fuels and somatic embryogenesis can improve plant breeding for this purpose. Sugarcane somatic embryo development involves
DOI:10.1093/jxb/ery209URLPMID:29860272 [本文引用: 1]
CRY1 interacts with HBI1 in a blue light-dependent manner. CRY1 and HBI1 antagonistically regulate expression of genes promoting cell elongation and CRY1 represses HBI1’s transcriptional activity by inhibiting its DNA binding ability. Cryptochromes (CRYs) are blue light photoreceptors that mediate various light responses in plants and animals. In Arabidopsis, there are two homologous CRYs, CRY1 and CRY2, which mediate blue light inhibition of hypocotyl elongation. It is known that CRY2 interacts with CIB1, a basic helix–loop–helix (bHLH) transcriptional factor, to regulate transcription and floral induction. In this study, we performed yeast two-hybrid screening and identified CIB1 as a CRY1-interacting protein. Moreover, we demonstrated that CRY1 physically interacted with the close homolog of CIB1, HBI1, which is known to act downstream of brassinosteroid (BR) and gibberellin acid (GA) signaling pathways to promote hypocotyl elongation, in a blue light-dependent manner. Transgenic and genetic interaction studies showed that overexpression of HBI1 resulted in enhanced hypocotyl elongation under blue light and that HBI1 acted downstream of CRYs to promote hypocotyl elongation. Genome-wide gene expression analysis indicated that CRYs and HBI1 antagonistically regulated the expression of genes involved in regulating cell elongation. Moreover, we demonstrated that CRY1–HBI1 interaction led to inhibition of HBI1’s DNA binding activity and its target gene expression. Together, our results suggest that HBI1 acts as a new CRY1-interacting protein and that the signaling mechanism of CRY1 involves repression of HBI1 transcriptional activity by direct CRY1–HBI1 interaction.
DOI:10.1073/pnas.1607687113URLPMID:27325768 [本文引用: 1]
BBX21 (also known as SALT TOLERANCE HOMOLOG 2), a B-box (BBX)-containing protein, has been previously identified as a positive regulator of light signaling; however, the precise role of BBX21 in regulating seedling photomorphogenesis remains largely unclear. In this study, we report that CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1) interacts with BBX21 in vivo and is able to ubiquitinate BBX21 in vitro. Thus, BBX21 is targeted for 26S proteasome-mediated degradation in dark-grown Arabidopsis seedlings in a COP1-dependent manner. Moreover, we show that BBX21 binds to the T/G-box in the ELONGATED HYPOCOTYL 5 (HY5) promoter and directly activates HY5 expression in the light. Transgenic seedlings overexpressing BBX21 exhibit dramatically shortened hypocotyls in the light, and this phenotype is dependent on a functional HY5. Taken together, our data suggest a molecular base underlying BBX21-mediated seedling photomorphogenesis, indicating that BBX21 is a pivotal component involved in the COP1-HY5 regulatory hub.
DOI:10.1104/pp.16.01067URLPMID:27784768 [本文引用: 1]
Abstract De novo organogenesis, which gives rise to adventitious roots and shoots, is a type of plant regeneration for survival after wounding. In Arabidopsis thaliana, two main cell fate transition steps are required to establish the root primordium during de novo root organogenesis from leaf explants. The first step from regeneration-competent cells to root founder cells involves activation of WUSCHEL-RELATED HOMEOBOX 11 (WOX11) and WOX12 (WOX11/12) expression by auxin. However, the molecular mechanism controlling the second step of fate transition from root founder cells to root primordium is poorly understood. In this study, we show that the expression levels of WOX11/12 decrease while those of WOX5 and 7 (WOX5/7) increase during the transition from root founder cells to the root primordium. WOX11/12 function genetically upstream of WOX5/7, and the WOX11/12 proteins directly bind to the promoters of WOX5/7 to activate their transcription. Mutations in WOX5/7 result in defective primordium formation. Overall, our data indicate that the expression switch from WOX11/12 to WOX5/7 is critical for initiation of the root primordium during de novo root organogenesis. {copyright, serif} 2016 American Society of Plant Biologists. All rights reserved.
DOI:10.3969/j.issn.1003-4315.2010.01.010URL [本文引用: 1]
采用发光二级管（LED）调制光质和光量,研究不同光质处理对叶用莴苣品种‘奶油生菜’和‘美国大速生’幼苗叶片气体交换参数及叶绿素荧光的影响.结果表明：LED光源光质提高了叶绿素a、叶绿素b、叶绿素总含量及叶绿素a/b值,品种间叶绿素含量差异显著,‘奶油生菜’〉‘美国大速生’;光质对叶用莴苣幼苗叶片光合速率的影响是由非气孔因素引起的,蓝光LED增大了叶片Gs,‘奶油生菜’以蓝光LED的光合速率最高,而‘美国大速生’以荧光灯处理最高;LED-红∶蓝下叶用莴苣各品种均具有最高的原初光能转化效率、开放的PSII中心有效光化学转化效率、PSII的电子传递速率和叶片PSII潜在活性;与荧光灯相比,LED光源对提高叶用莴苣叶片光合能力具有明显优势,有利于提高叶片的PSII活性和QA的还原速率.
DOI:10.3969/j.issn.1003-4315.2010.01.010URL [本文引用: 1]
采用发光二级管（LED）调制光质和光量,研究不同光质处理对叶用莴苣品种‘奶油生菜’和‘美国大速生’幼苗叶片气体交换参数及叶绿素荧光的影响.结果表明：LED光源光质提高了叶绿素a、叶绿素b、叶绿素总含量及叶绿素a/b值,品种间叶绿素含量差异显著,‘奶油生菜’〉‘美国大速生’;光质对叶用莴苣幼苗叶片光合速率的影响是由非气孔因素引起的,蓝光LED增大了叶片Gs,‘奶油生菜’以蓝光LED的光合速率最高,而‘美国大速生’以荧光灯处理最高;LED-红∶蓝下叶用莴苣各品种均具有最高的原初光能转化效率、开放的PSII中心有效光化学转化效率、PSII的电子传递速率和叶片PSII潜在活性;与荧光灯相比,LED光源对提高叶用莴苣叶片光合能力具有明显优势,有利于提高叶片的PSII活性和QA的还原速率.
DOI:10.3389/fpls.2017.00857URLPMID:5440764 [本文引用: 1]
The aim of this study was to investigate the effects of light quality on the morphological traits, leaf anatomical characteristics, antioxidant enzyme (superoxide dismutase, catalase, and peroxidase) activities, photosynthetic pigments content, and bioactive compounds (phenols, flavonoids, and polysaccharides) content inAnoectochilus roxburghii. Plants ofA. roxburghiiwere grown under light filtered through four differently colored films for 8 months. The four treatments were red film (RF), blue film (BF), yellow film (YF), and colorless plastic film (control, CK). Compared with theA. roxburghiiplants in CK, those in the BF treatment showed significantly greater stem diameter, fresh weight, leaf area, stomatal frequency, chlorophyll content (Chl a, Chl b, Chl a+b), antioxidant enzyme activities, and active compound (polysaccharides, flavones) content. The plants in the RF treatment showed the greatest plant height and phenolics contents. These results show that growingA. roxburghiiplants under blue film is a useful technique to improve quality. This technique is conducive to achieving large-scale sustainable production of high-quality plant materials.
