长期CO_2加富对苗期红掌(Anthurium andraeanum L.)植株生长和光合作用的影响

以开顶式塑料薄膜温室为设施,研究了红掌(AnthuriumandraeanumL.)幼苗植株生长、叶片净光合速率和光合酶活性对长期高CO2浓度的响应。结果表明:处理90d时,处理组T1((700±100)μmol·mol-1CO2)的株高、单叶面积、株鲜重分别比对照组((360±30)μmol·mol-1)增加了15.76%、14.30%、29.62%,而处理组T2((1000±100)μmo·lmol-1CO2)的株高、单叶面积、株鲜重分别比对照增加了15.00%、9.63%、36·22%;处理150d时T1的株高、单叶面积、株鲜重与对照相比分别增加了16·08%、17.30%、49.09%,而T2增加了16.61%、10.10%、48.87%。在各自生长环境下处理组T1、T2的净光合速率在整个处理期间均高于对照,处理150d时,T1、T2的净光合速率分别比对照高8.25%、20.62%;但处理90d时,在对照CO2浓度下测定的净光合速率处理组开始低于对照组,可能此时处理组的红掌叶片开始出现光合适应现象;CO2浓度升高促进了叶片中可溶性糖和淀粉积累,处理90d时T1、T2处理组中淀粉含量分别比对照高52.60%、67.66%;处理150d时,T1组红掌叶片中淀粉与可溶性糖含量比对照高53.43%、6.32%,T2比对照高58.44%、8.07%,叶绿素含量在处理90d时也开始低于对照组;整个实验过程中,Rubisco活性前期增加,90d以后开始下降;乙醇酸氧化酶活性则明显下降,T1、T2处理组试验结束时与对照组相比分别下降了41.28%、45.35%。一定处理时间(90d)的高浓度CO2处理提高了红掌叶片的净光合速率和碳水化合物的积累,促进了营养生长,但随着处理时间的延长,这种促进作用逐渐降低。

Plant Regeneration by Callus-Mediated Protocorm-Like Body Induction of Anthurium andraeanum Hort.

This research aims at developing a plant regeneration system from leaf and petiole explants of Anthurium andraeanum Hort., thereby establish a foundation for mass production and transformation. Using tissue culture technique, the conditions for callus induction, protocorm-like body （PLB） formation and plant regeneration from leaf explants and petiole of A. andraeanum, such as basal medium and plant growth regulator, were investigated. Totipotent callus was induced on a 1/2-strength MS medium containing 0.90 μmol L^-1 2,4-dichlorophenoxyacetic acid （2,4-D） and 8.88μmol L^-1 N6-benzyladenine （BA）. The callus exhibited complete hormone autonomy for growth and differentiation of PLBs. This callus proliferated well and was maintained by subculturing on 1/2 MS medium containing 0.90 μmol L^-1 2,4-D and 4.44 μmol L^-1 BA. On average, 8 protocorm-like bodies could be obtained from a piece of 4 mm callus after being transferred to the 1/2 MS medium with 4.44 μmol L^-1 BA after 8 wk of culture. The regenerated PLBs formed shoots and roots on 1/2 MS medium. After 24 wk of culture on these medium, well-developed plantlets for potting were produced. An efficient micropropagation method was established for indirect PLB formation and plant regeneration from leaf and petiole ofA. andraeanum.

Advances in Transgenic Breeding of Anthurium andraeanum

At present, transgenic technologies have become important means of plant breeding, and the application and promotion of transgenic technologies have created huge economic and social benefits. Transgenic plant products have significantly affected human life. Anthurium andraeanum is the second major tropical potted flower and its transgenic breeding has a promising prospect of application. In this paper, acceptors, transformation methods and introduced exogenous genes （ including reporter genes, selectable marker genes and target genes） of Anthurium andraeanum were summarized; in addition, several issues related to transforma- tion of Anthurium andraeanum were analyzed, aiming at providing reference for transgenic breeding of Anthurium andraeanum.

1-DE/MS Analysis of Proteins Related to the Spathe Color Variation in an Anthurium andraeanum Mutant

[Objectives] This study was conducted to explore the color variation mechanism of Anthurium andraeanum spathe at the protein level. [Methods]The differential proteins of wild type and its white mutant were separated and identified by using one-dimensional gel electrophoresis and mass spectrometry( 1-DE/MS). [Results] Compared with leaves and spadices,the 1-DE patterns of two kinds of spathe proteins were significantly different,and two different bands were detected in wild type spathes and mutant spathes respectively. The four significantly differential bands were selected and analyzed by mass spectrometry,and 138,111,70 and 427 proteins were identified respectively. The results of GO functional annotation analysis showed that the molecular functions of the proteins were mainly catalytic activity and binding,and the main biological processes involved were cellular process and metabolic process. Many proteins involved in the synthesis of anthocyanins and flavonoids,sugar metabolism and some resistance proteins were screened,indicating that the spathe color difference of A. andraeanum‘Pink champion'is not only related to anthocyanin anabolism,but also regulated by various metabolic pathways. [Conclusions]The study provides a new experimental basis for elucidating the molecular mechanism of the regulation of A. andraeanum flower color.

1-DE/MS Analysis of the Proteins Related to Spathe Color Variation in Anthurium andraeanum 'Madural'

In order to understand the mechanism of spathe color variation in Anthurium andraeanum at the protein level, the leaves, inflorescences and spathes of the wild type and two mutants of A. andraeanum 'Madural' were used as research objects in which the differential expression of proteins related to flower color mutants was analyzed by one-dimensional electrophoresis and mass spectrometry(1-DE/MS). The 1-DE patterns showed that the protein components expressed highly in spathes were mainly concentrated in the molecular weight range of 20-42 kD, and differential bands were detected between the wild type and the mutant, while no significant differences were detected in the leaf and inflorescence proteins. According to the results of mass spectrometry analysis of the differential bands, 21 known functional proteins involved in life processes such as glucose metabolism, resistance, cytoskeleton, gene regulation and signal transduction were identified. It showed that in addition to the influences from anthocyanidins, the spathe color variation of A. andraeanum 'Madural' is also regulated by a variety of metabolic pathway-related proteins.

红掌(Anthurium andraeanum)细胞工程研究进展及其在育种上的应用

红掌是一种重要的观花和观叶兼具的草本植物。植物离体培养技术已经成为红掌繁殖主要采用的方法,绝大部分品种可以通过组织培养实现再生,其推动了红掌细胞工程育种的发展。本综述总结了红掌细胞组织培养繁殖途径,包括直接获得无菌苗途径、愈伤组织诱导和不定芽再生途径、体细胞胚胎发生途径、原球茎和类原球茎途径、原生质体途径等方面的研究进展,及其花器官培养,多倍体诱导,物理化学诱变,遗传转化,体细胞无性系变异及筛选的研究在育种中的应用。并提出了红掌细胞工程研究当前存在的问题和以后的研究方向,以期为将来红掌的研究提供借鉴。

Cloning and functional analysis of a novel ascorbate peroxidase(APX) gene from Anthurium andraeanum

An 888-bp full-length ascorbate peroxidase(APX) complementary DNA(cDNA) gene was cloned from Anthurium andraeanum, and designated as AnAPX. It contains a 110-bp 5′-noncoding region, a 28-bp 3′-noncoding region, and a 750-bp open reading frame(ORF). This protein is hydrophilic with an aliphatic index of 81.64 and its structure consisting of α-helixes, β-turns, and random coils. The AnAPX protein showed 93%, 87%, 87%, 87%, and 86% similarities to the APX homologs from Zantedeschia aethiopica, Vitis pseudoreticulata, Gossypium hirsutum, Elaeis guineensis, and Zea mays, respectively. AnAPX gene transcript was measured non-significantly in roots, stems, leaves, spathes, and spadices by real-time polymerase chain reaction(RT-PCR) analysis. Interestingly, this gene expression was remarkably up-regulated in response to a cold stress under 6 °C, implying that AnAPX might play an important role in A. andraeanum tolerance to cold stress. To confirm this function we overexpressed AnAPX in tobacco plants by transformation with an AnAPX expression construct driven by CaMV 35S promoter. The transformed tobacco seedlings under 4 °C showed less electrolyte leakage(EL) and malondialdehyde(MDA) content than the control. The content of MDA was correlated with chilling tolerance in these transgenic plants. These results show that AnAPX can prevent the chilling challenged plant from cell membrane damage and ultimately enhance the plant cold tolerance.

粉掌盆花模拟贮运保鲜效果研究

以粉掌盆花为试材,进行水杨酸(SA)、硫代硫酸银(STS)和1-甲基环丙烯(1-MCP)3种处理,通过测定粉掌盆花叶片的相对电导率、叶绿素荧光参数、丙二醛(MDA)含量、相对含水量等生理指标,研究模拟贮运黑暗条件下3种处理对粉掌盆花的保鲜效果。结果表明,随着处理天数的增加,经SA、STS、1-MCP处理的粉掌叶片相对电导率均有所增加,叶绿素荧光参数F_(o)、F_(m)和F_(v)/F_(m)总体上均呈下降的趋势,MDA含量则不断增加,相对含水量减少。与对照相比,SA、STS和1-MCP均有利于延长粉掌盆花的寿命,其中1-MCP处理对粉掌盆花的保鲜效果最好。

基于层次分析法的红掌种质资源评价

红掌是重要的盆栽、切花均可的叶花兼用型花卉,品种众多、形态各异,具有较高的观赏价值。通过对157个红掌品种的36个性状进行观测,依据各性状的表达状态、分布频率及对观赏性的影响,建立一套评分标准,并采用层次分析法对红掌的种质资源进行综合评价。结果表明:佛焰苞大小的权重系数最大,达到0.114;花梗长度、佛焰苞上表面主色、佛焰苞下表面主色、佛焰苞光泽度、佛焰苞泡状程度、肉穗花序花粉囊即将开裂时基部的主色、肉穗花序花粉囊即将开裂先端的主色7个性状的权重系数均大于0.05;157个品种分值为2.536-6.100,其中‘香妃’‘安祖伊维斯’和‘安祖亚柯索’3个品种综合得分均大于6,中等大小的红色、粉色品种得分最低,而超大型品种或迷你品种比较少见。

红掌小滴玻璃化法超低温保存研究

以红掌腋芽为试材,探讨小滴玻璃化法对红掌超低温保存的影响因素,并对再生植株进行遗传稳定性检测。结果表明,红掌腋芽置于含0.4 mmol/L蔗糖和2 mmol/L甘油的MS固体培养基中预培养4 d后,在0℃下80%PVS_(2)处理50 min,转到铝箔条上PVS_(2)液滴中,将铝箔条迅速浸入液氮中,2 s后直接转入装满液氮的冻存管中,投入液氮至少保持30 min;室温下用含有1.2 mmol/L蔗糖的MS液体培养基复温并卸载20 min后置于恢复培养基上,腋芽存活率高达63.70%。通过ISSR和SSR分子标记检测,再生植株的遗传稳定性未发生改变。该研究结果为红掌种质资源的长期保存提供有效途径。

红掌新品种‘阳光红心’组织培养与快繁技术研究

【目的】建立红掌新品种“阳光红心”种苗繁育技术体系。【方法】以阳光红心为试验材料,开展不同外植体、激素组合对其组织培养过程中诱导、增殖和生根的影响。【结果】适宜建立阳光红心无菌系的外植体是叶片,初代诱导培养基为改良MS+1.0mg/L6-BA+0.2mg/LNAA,最高诱导率为56.5%;最佳继代培养基组合为改良MS+2.0mg/LBA+0.5mg/LNAA+0.5mg/LKT,繁殖系数达5.8;最佳的生根培养基为1/2MS+0.5mg/LNAA,生根率达100%;泥碳土:珍珠岩:椰糠=5:3:2的基质组合最有利红掌的移栽生长,移栽成活率为94.8%。【结论】建立阳光红心组织培养快繁技术体系,为红掌植物的规模化繁殖提供理论依据。

广东地区红掌橘全爪螨危害调查

通过显微观察橘全爪螨生活史与产卵量,开展广东地区红掌橘全爪螨危害普查和调查,结果发现橘全爪螨从若螨发育为雌成螨的当天不产卵,第2天开始每天产卵3~4枚,多则5枚,产卵历时8 d,每雌螨总产卵量约28.40枚。雌螨生命周期约为17.90 d,从卵期、幼螨期、若螨期到成螨期分别为5.56、1.42、2.52和8.40 d。橘全爪螨主要危害红掌中下部老叶,以危害叶背为主,虫害发生初期叶背出现许多灰白色小点,严重时全叶呈灰白色。不同红掌品种橘全爪螨的危害虫情指数差异很大,‘维多’红掌受害最重,虫情指数20.4%;‘朝天娇’‘广花福运’‘红斑比诺’‘满天星’‘莉莉’5个红掌品种受害较轻,虫情指数均小于5.0%。本研究为提前预防螨虫危害和开展有效药剂筛选提供了科学依据。

红掌四倍体的离体诱导及其鉴定

以愈伤组织为诱导材料,采用秋水仙素处理方法进行了红掌四倍体的离体诱导。结果发现,红掌四倍体诱导率因处理方法、秋水仙素浓度和处理时间不同而异。采用液体培养的四倍体诱导率比固体培养的高,低浓度秋水仙素的诱导率比高浓度的高,长时间处理的诱导率比短时间处理的高,基因型对四倍体诱导率影响不明显。在含0·2g·L-1秋水仙素的液体培养基中振荡培养14d,四倍体诱导率最高,为45·5%。四倍体红掌的花药和花粉比二倍体的大,植株粗壮,叶片和佛焰苞大而厚,颜色深,叶片下表皮上的气孔密度小,长度长。改良压片法与气孔长度和密度鉴定法是鉴定红掌四倍体的可靠方法。

红掌气生根根段再生快繁体系的建立

以红掌气生根根段为材料,诱导再生团块产生,进而分化出苗,形成快速繁殖系统。培养基1/2MS+6-BA1.0mg·L-1+2,4-D0.6mg·L-1适于气生根的保持和繁殖,1/2MS+6-BA1.0mg·L-1+2,4-D0.2mg·L-1适于诱导气生根再生团块的产生,MS+6-BA1.0mg·L-1+2,4-D0.2mg·L-1可使再生团块分化成苗。不论是愈伤组织,还是再生团块,出现绿色组织是分化所必需的。添加2,4-D、6-BA和2,4-D的适当比例、MS培养基的无机盐浓度在再生团块的诱导与分化成苗中起重要作用。

红掌愈伤组织诱导和芽的分化

对影响红掌愈伤组织诱导及芽分化的几个因素进行了研究。同一成熟度的外植体 ,其叶柄愈伤组织诱导率、芽分化率、分化时间均明显优于叶片。不同放置方式和光照时间对叶片愈伤组织的诱导亦有影响 ,叶背向下放置 ,光照时间 2 4h/d和 10h/d愈伤组织诱导率较高 ,分别为 10 0 %、 97%。光照时间对叶柄愈伤组织诱导无显著影响 ,但光照 2 4h/d和 10h/d较无光照处理的明显促进芽的分化。叶柄培养以N6,KC和 1/2MS培养基为佳。叶片培养则以P ,N6和 1/2MS为好。以未展叶叶柄为外植体 ,从接种到丛芽分化共 4 9d ,较已有报道提前 11～ 31d。

LED光源不同光质比例对红掌试管苗生长的影响

以红掌品种"骄阳"试管苗为材料,研究LED光源不同光质比例对红掌试管苗生长的影响。结果表明:在70%R(红光)+30%B(蓝光)处理下,红掌试管苗的株高、叶数、干鲜质量、根系活力达到最大值;在80%R+20%B处理下,红掌试管苗的根数、根长、可溶性糖含量达到最大值;在50%R+50%B处理下,红掌试管苗的叶长、叶幅达到最大值,且在此处理下叶绿素a、叶绿素a+b、叶绿素a/b均达到最大值;蓝光处理下有利于干物质积累。红蓝组合光处理的红掌试管苗较单一红蓝光有明显优势,在50%R+50%B处理下有利于红掌试管苗的形态建成,其根系活力、可溶性糖含量均显著高于对照,是适宜红掌试管苗生长的最佳红蓝光配比。试验结果表明LED光源对促进红掌试管苗的生长和提高其品质效果明显。

高温对红掌幼苗叶片茉莉酸浓度及抗氧化系统的影响

试验研究30℃和38℃高温处理对红掌幼苗叶片茉莉酸(JAs)浓度及抗氧化系统的影响,结果表明:38℃高温处理6h,叶片内JAs浓度急剧升高,其后又呈下降趋势,但其浓度仍高于对照;30℃高温处理12h,叶片内JAs浓度达最大值,之后随处理时间的延长而降低,但仍高于对照。高温处理后叶片抗氧化系统中的过氧化物酶(POD)、超氧化物歧化酶(SOD)活性均有不同程度升高,O 2含量在高温处理的前期有所降低,后期又逐渐升高。一定程度的高温处理可通过提高JAs浓度和抗氧化酶活性来提高红掌幼苗的抗热性。

花烛体细胞胚胎发生及植株再生研究

以花烛(Anthurium andraeanumLind.)盆花品种无菌苗叶片、叶柄、根段为外植体,对花烛体细胞胚胎发生及植株再生进行了研究,建立了花烛体细胞胚胎发生体系。结果表明,花烛叶片较其他外植体更适宜体细胞胚的诱导;胚性愈伤组织及胚性结构诱导最适培养基为改良MS+2,4-D2.0～3.0mg.L-1+KT0.5mg.L-1+蔗糖2%+葡萄糖1%,pH5.5,暗培养40d,诱导率可达90.3%;体胚发育适宜培养基为改良MS+6-BA0.5mg.L-1+蔗糖3%,光强20μmol.m-2.s-1,培养20d后体胚萌发率达62.9%;萌发的体胚在发育培养基上继续培养30d后均能发育成完整植株。对不同阶段培养材料的组织结构及超微结构的观察证明了花烛体胚发生过程。

CO_2浓度升高对红掌光合速率与生长发育的影响

本试验以开顶式塑料薄膜温室为设施,研究了高CO2浓度对红掌叶片光合速率、植株生长、光合酶活性和花期的影响。结果表明:处理90d时,对照组〔大气CO2浓度(360±30)μmol·mol-1〕红掌的株高、叶面积、株干样质量、株鲜样质量与处理前相比分别增加了16.4%、36.1%、101.2%和84.2%,而高CO2浓度组〔(1000±100)μmol·mol-1〕则分别增加了72.9%、65.6%、217.6%和199.1%。高CO2浓度组的净光合速率比对照增加46.27%,气孔导度下降,促进了叶片中可溶性糖和淀粉积累,叶绿素含量比对照下降,而对Rubisco活性影响较小,乙醇酸氧化酶活性则明显下降。高CO2浓度处理50d时,开花率为25%,处理90d时已达到80%以上,而在整个试验期间对照组未见开花。因此,高浓度CO2处理提高了红掌叶片的光合速率和碳水化合物的积累,促进了营养生长,提前了花期。

园林废弃物堆肥替代泥炭用于红掌和鸟巢蕨栽培

为促进园林废弃物资源化利用,将改良后的园林废弃物堆肥分别以0%,20%,40%,60%,80%,100%替代泥炭进行红掌Anthurium anaraeanum和鸟巢蕨Asplenium nidus栽培研究。该研究共设6个处理3次重复。结果表明：园林废弃物堆肥的添加显著提高栽培基质容重、持水孔隙,降低了总孔隙度、通气孔隙（P〈0.05）;园林废弃物堆肥的添加显著降低基质有机碳质量分数（P〈0.05）,提高基质p H值、电导率值、营养元素质量分数（P〈0.05）;鸟巢蕨栽培中,以添加园林废弃物堆肥60%~80%效果明显,植株鲜质量、株高、冠幅、叶绿素质量分数分别提高11.91%~51.18%,4.09%~5.12%,31.72%~43.08%,18.06%~14.84%（P〈0.05）;红掌栽培中,以添加园林废弃物堆肥60%效果最好,鲜质量、株高、冠幅,叶绿素质量分数分别提高27.00%,23.74%,13.05%,10.15%（P〈0.05）。园林废弃物堆肥可以部分替代泥炭用于红掌和鸟巢蕨栽培。