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。

Differences in Primary Callus Induction among Different Anthurium andraeanum Varieties

[Objective] This research aimed to optimize continuously the highly efficient regeneration system of Anthurium andraeanum. [Method] The leaves and petioles of four A. andraeanum varieties were used as explants to investigate the differences in primary callus induction among different A. andraeanum varieties. [Result]The callus formation capacity of SAM and SST was stronger than that of SDM and SHG. Among the four varieties, the leaf regeneration capacity of SAM, SDM and SHG was stronger than the corresponding petiole regeneration capacity. However,the petiole regeneration capacity of SST was stronger. The optimum medium for petiole callus induction of SST was 1/2 MS ＋ TDZ 4.0 mg/L ＋ 2, 4-D 0.2 mg/L with induction rate of 87.5%; the optimum medium for leaf callus induction of SAM was 1/2 MS ＋ TDZ 2.0 mg/L ＋ 2, 4-D 0.2 mg/L with induction rate more than90%; the optimum medium for leaf callus induction of SDM and SHG was all 1/2MS ＋ ZT 2.0 mg/L ＋ 2, 4-D 0.2 mg/L with induction rates of 59.34% and 79.63%,respectively. [Conclusion] In addition to variety differences, the differences in differentiation ability among different types of calluses should be also taken into account in the establishment and optimization of tissue culture and rapid propagation technology system of A. andraeanum.

Key Techniques of Industrial Seedling for Anthurium andraeanum

[Objective] The aim was to study the techniques for the tissue culture,rapid propagation and transplantation of Anthurium andraeanum.[Method] The leaf,leaf stalk,spathe and spadix of the pot flower variety and cut flower variety of A.andraeanum were used as explants,in order to investigate the influences of the explants selection,disinfection conditions and different media on the callus induction,proliferation and rooting.[Result] The leaf stalk of A.andraeanum was the optimum explant,with the highest callus induction rate（78.3%）.The best disinfecting effect could be obtained when the explants was sterilized with 0.1% HgCl2 for 8 min.The optimum media for the callus induction of Ping Champion and MIDORI were respectively 1/2 MS＋1.50 mg/L 6-BA＋0.50 mg/L 2,4-D＋0.10 mg/L NAA and 1/2 MS＋1.50 mg/L 6-BA＋1.00 mg/L KT＋0.10 mg/L 2,4-D,while the optimum media for callus subculture of Ping Champion and MIDORI were respectively MS＋1.00 mg/L 6-BA＋0.50 mg/L NAA＋0.30 mg/L 2,4-D and MS＋0.50 mg/L 6-BA＋0.50 mg/L KT＋0.10 mg/L NAA.It was beneficial for the proliferation and growth of cluster buds while 10%（w/v）of coconut water or banana jam was added to MS medium at the proliferation stage,and the proliferation rate could be increased by 2.83 fold.At the rooting stage,the medium 1/2 MS＋0.10 mg/L NAA＋0.10 mg/L IAA was the optimum medium for rooting,and the rooting rate was up to 100%.[Conservation] The research results will lay a foundation for the establishment of the industrial seedling breeding system of A.andraeanum,and it will be significant for the development of A.andraeanum industry.

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 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.

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.

硼及基质对红掌“Dakota”生理特性的影响

以红掌"火焰"为试材,研究硼及基质对植株生理特性的影响,并筛选较优的处理组合。试验结果表明:同种基质中,以硼40μmol/L时根系活力较高,而叶绿素含量却在20μmol/L时较高。SOD和POD活性在不同基质中变化规律不同;而MDA含量在不同基质中变化规律相同,以硼20μmol/L时最高。试验范围内,硼及基质处理间的互作效应明显,且以A4B2处理组合较优。

红掌特伦萨四倍体的离体诱导及其鉴定

为了研究红掌四倍体的离体诱导和鉴定技术体系,为其倍性育种和杂交育种奠定基础,以特伦萨愈伤组织为试验材料,采用0.5、1.0、2.0、4.0 g·L^(-1)的秋水仙素分别处理2、4、6、8 h,之后转入培养基MS+6-BA 2.0 mg·L^(-1)+NAA 0.5 mg·L^(-1)进行愈伤组织的生长和丛生芽分化,75 d后将丛生芽转入培养基1/2 MS+NAA 0.2 mg·L^(-1)诱导生根,待丛生芽生根后统计愈伤组织块存活率、再生植株数,并对再生植株进行染色体倍性鉴定和核型分析。结果表明,随着秋水仙素浓度的增加,红掌愈伤组织块存活率呈下降趋势,但愈伤组织块上诱导的再生植株数无显著变化;在0.5~2.0 g·L^(-1)的秋水仙素浓度范围内,红掌疑似四倍体的诱导率随着处理浓度与时间的增加而增加,2.0 g·L^(-1)秋水仙素处理6与8 h时,诱导率达8.50%与8.77%;特伦萨和疑似四倍体红掌的基因组分别为3.73和7.01 Gb;染色体压片倍性鉴定结果证明了疑似四倍体的倍性为四倍体。红掌二倍体的核型公式为2n=2x=30=8 st+16 sm+6 m,四倍体的核型公式为2n=4x=60=8 st+32 sm+20 m;与红掌二倍体植株相比,四倍体植株叶片变厚、气孔变大、保卫细胞密度变小,佛焰苞直径变大,单个花序花期变长,差异均达显著水平。综上,采用2.0 g·L^(-1)秋水仙素处理6 h可培育红掌四倍体植株,此四倍体可作为红掌倍性育种和杂交育种的重要种质资源。

‘Dakota’红掌营养诊断采样部位的研究

通过对红掌品种‘Dakota’各部位N、P、K和Ca含量与平衡比例的分析,确定了一年生和二年生植株大量元素诊断的采样部位分别为新近成熟叶片与新叶片。对红掌各部位营养元素分布规律研究表明,可通过一定系数将营养诊断测定结果统一为全株含量,消除各部位与全株间元素含量存在的差异。一年生植株的新近成熟叶片N、P、K和Ca系数分别为1.828、1.689、1.437和2.505,二年生的为1.339、1.282、1.282和0.676。

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。

红掌组织培养光调控技术研究进展

红掌是国内外花卉市场颇受欢迎的重要植物之一,具有较高的经济价值和观赏价值,在国内外花卉研发和市场应用方面具有广阔的前景。植物组织培养技术繁育质量高稳,繁育效率高,不受外界气候、地域、时间等诸多因素的制约。植物组织培养技术成为红掌种质资源离体保存与繁育、种质资源创制和工厂化育苗的重要手段。光照条件是影响红掌组织培养的核心因素之一。基于此,综述了红掌组织培养光调控技术研究进展,总结和分析了光条件对红掌愈伤组织诱导、增殖培养、组培苗生长等的影响,提出了相关展望。

利用CTAB法提取红掌不同部位基因组DNA

采用CTAB法,对红掌的叶、叶柄、肉穗花序、佛焰苞、根等5个部位进行基因组DNA提取,利用紫外分光光度法、琼脂糖凝胶电泳法和RAPD扩增法检测DNA的质量,结果表明,CTAB法适合红掌根和叶的基因组DNA提取,没有蛋白质、酚及色素等杂质的污染,佛焰苞的DNA中有少量蛋白质污染,叶柄DNA中含有较多的蛋白质、酚等杂质,而从肉穗花序中没有提取到基因组DNA。

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。

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。

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。

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。

红掌种质资源形态学标记与遗传多样性分析

为了解红掌种质资源的遗传基础,对收集的32个红掌品种的形态学性状如株高、花色、叶色等进行比较分析。基于形态学标记,这些品种的遗传多样性丰富。UPGMA聚类分析表明,这32个红掌品种可划分为5大类,其中包括4个较小的类群和1个较大的类群。本研究鉴定的一些形态学标记对于红掌品种鉴定、遗传多样性分析和杂交育种具有重要的应用价值。