五唇兰对PEG模拟的干旱胁迫响应研究

为了解干旱对五唇兰(Phalaenopsis pulcherrima)生长的影响,以聚乙二醇(PEG)溶液模拟干旱胁迫,对其叶片的光合色素、渗透调节物质和非结构碳水化合物(NSC)含量变化进行研究。结果表明,随着PEG浓度增加,五唇兰植株含水量和鲜质量逐渐下降,以PEG为13.75%～14.84%时最显著。PEG处理显著降低叶片的叶绿素a和b含量。随着植株含水量的降低,叶片可溶性蛋白、淀粉(St)含量均呈下降趋势,可溶性糖(SS)含量、NSC和SS/St均呈先升后降的趋势。因此,干旱胁迫会影响五唇兰植株的含水量和光合产物的积累;在较低程度干旱胁迫下,可溶性糖在抗旱响应中发挥主要作用;随着干旱胁迫程度加深,五唇兰的生理代谢受到严重影响。     

PEG胁迫及复水对不同抗旱性小麦幼苗脯氨酸代谢关键酶活性的影响

以两种不同抗旱性小麦品种幼苗为试验材料,采用PEG模拟干旱胁迫处理,探究干旱胁迫及复水对小麦幼苗叶片与根系脯氨酸累积及关键酶活性的影响。结果显示:(1)PEG胁迫下抗旱品种‘普冰143’根长和根干重下降不大,而水敏感品种‘郑引1号’根长和根干重下降显著;且于胁迫处理36h时‘普冰143’根系脯氨酸含量增加(75.0%)显著大于‘郑引1号’(37.7%),复水24h后均恢复至对照水平。(2)PEG胁迫下‘普冰143’叶片中谷氨酸合成途径关键酶P5CS和鸟氨酸合成途径关键酶δ-OAT活性均显著增加,且‘普冰143’叶片脯氨酸两条合成途径关键酶活性均得以加强;PEG胁迫处理36h时,‘郑引1号’叶片中P5CS活性增加显著,δ-OAT活性变化较小,且‘郑引1号’叶片脯氨酸合成可能以谷氨酸途径为主;但在PEG胁迫下两个不同抗旱性品种的根中P5CS、δ-OAT活性均变化较小。(3)PEG胁迫处理36h时‘普冰143’叶片脯氨酸降解酶PDH活性显著下降,而‘郑引1号’叶片PDH活性显著增加,复水后抗旱品种叶片该酶活性显著增加,水敏感品种恢复至对照水平;但PEG胁迫处理下两个不同抗旱性品种的根中PDH活性均显著下降。研究表明,PEG胁迫下小麦叶片是合成脯氨酸的主要部位,抗旱品种‘普冰143’根系脯氨酸持续积累与叶片中高的脯氨酸合成关键酶活性及脯氨酸转运有关。     

等渗的盐分和水分胁迫对芦荟幼苗生长和离子分布的效应

 研究了等渗透势（-0.44、-0.88 MPa）NaCl和PEG 6000处理对六叶龄芦荟(Aloe vera)幼苗叶片生长速率、干物质积累、电解质渗漏和离子吸收、分配的效应。结果表明: -0.44、-0.88 MPa NaCl和PEG处理10 d均明显抑制芦荟幼苗叶片伸长生长,植株干物质积累速率显著降低, 叶片含水量降低,叶片细胞电解质渗漏率上升。NaCl对芦荟幼苗生长的抑制作用显著大于PEG处理的。不同器官离子含量、根系和叶片横切面X-射线微区分析结果表明, NaCl胁迫导致芦荟体内Na+、Cl－含量显著上升,根中增幅明显高于叶片,其中Cl－尤为显著。NaCl胁迫严重抑制芦荟对K+ 和Ca2+ 的吸收及其向叶片的运输,根、叶K+/Na+、Ca2+/Na+ 比率显著下降,而PEG胁迫对离子平衡的干扰较轻,是芦荟对水分胁迫的适应能力高于盐胁迫的主要原因之一。但芦荟对 -0.44～-0.88 MPa NaCl胁迫仍有一定的适应能力,主要原因是：1) 根系对离子的选择性吸收和运输较强,并随着盐胁迫强度增加其选择性增强; 2) 芦荟叶片中的盐分在贮水组织中显著积累,明显高于其它组织细胞。同时,芦荟是CAM（景天酸代谢）途径植物,蒸腾极小,盐分随蒸腾流进入地上部的机会小。     

从胡萝卜与芹菜原生质体融合获得细胞杂种

试验以胡萝卜根和芹菜叶肉原生质体为亲本,用PEG高pH,高Ca~( )法诱导融合。异源融合率最高可达8％,一般在3％～5％之间。从150余棵融合再生植株中,经形态、同工酶谱、叶片中内含芳香物质等指标测试,鉴定出三棵(No 1,2,8)融合再生植株为芹莱和胡萝卜的细胞杂种。     

施硼对番木瓜幼龄株硼形态含量及叶片光合作用的影响

探讨根外喷硼与土壤施硼对植株硼形态含量及叶片光合作用的影响。结果表明,两种供硼方式均可增加植株叶片、叶柄及根的硼形态含量及硼总量,其中以叶片水溶态硼、半束缚态硼及硼总量的增加最为显著。植株硼含量增加提高了叶片净光合速率,但硼过量却抑制了叶片光合能力的提升,而硼中毒导致光合能力显著下降。     

供氮形态与部位对局部根区水分胁迫下玉米水分吸收与利用的影响

试验以玉米品种'金海五号'幼苗为材料,在分根条件下采用聚乙二醇(PEG-6000)模拟局部根区水分胁迫,设置3种供氮形态(硝态氮、铵态氮、两者各占50%的混合氮)和2种供氮部位(水氮同区,氮加入到无PEG侧;水氮异区,氮加入到含PEG侧),研究局部根区水分胁迫下氮形态与供应部位对玉米水分吸收和利用的调节与作用机制,为局部根区灌溉水分高效利用提供理论依据.结果发现:(1)同一氮形态下水氮同区供应的植株蒸腾速率、耗水量、木质部汁液流速和生物量较高,加有硝态氮源处理无PEG侧根系的导管数目及单一氮形态处理无PEG侧根系的导管直径较高,但木质部汁液、叶片中脱落酸(ABA)浓度以及水分利用效率均较低.(2)同一供氮部位下,植株的蒸腾速率、耗水量、木质部汁液流速和生物量的顺序均为混合氮>硝态氮>铵态氮依次,但单一铵态氮处理植株的ABA浓度较高,水分利用效率较高.研究表明,同一氮形态下水氮同区供应植株生长较好、水分吸收能力较强,但水氮异区供应下植株的水分利用效率较高;同一供氮部位下,植株生长和水分吸收能力的顺序为混合氮>硝态氮>铵态氮,但单供铵态氮植株的水分利用效率较高.     

辽东楤木皂苷分布特性的研究

目的:研究辽东楤木植株中的皂苷分布。方法:以1~8年株龄辽东楤木为试材,采用超声提取法提取楤木皂苷,采用分光光度计法测定辽东楤木植株的根和茎不同部位的皂苷含量,不同生长发育时期和不同光照条件下叶片的皂苷含量。结果:辽东楤木根皮、茎皮和叶片中均含有大量的皂苷成分,根皮和茎皮中的皂苷含量逐年增加,8年生植株的根、茎皮中分别为11.0%和8.2%,根、茎的木质部和髓中亦含有皂苷,但相对较少;叶片皂苷含量随株龄增长而增加,4、6年生植株叶片皂苷含量分别为8.3%、8.9%;光照处理对叶片的生长及皂苷含量影响较大,50%透光率及以下的光照条件叶片生长不良,早衰严重,皂苷积累较少,最高含量为5.9%,75%透光率条件同全光照条件叶片发育形态指标接近,并有利于皂苷形成,皂苷含量平均高于全光条件0.6%,结论:6年以上植株的根茎皮在皂苷含量和总量上都相对较高;4年以上植株成熟的叶片在皂苷含量和总量上均较高,最适宜作为药源材料;叶片在一定遮阳条件下更有利于皂苷形成与积累。     

烟草毛状根多倍体诱导及其植株再生

为了提高烟草的烟碱含量,采用发根农杆菌遗传转化和人工染色体加倍技术,进行了烟草毛状根及其多倍体诱导、植株再生及其烟碱含量测定。结果表明,发根农杆菌ATCC15834感染烟草叶片外植体8 d后产生白色毛状根,15 d后所有叶片外植体产生毛状根。毛状根能在无外源激素的MS固体和液体培养基上自主生长。PCR扩增结果显示发根农杆菌Ri质粒的rol B和rol C基因以及冠瘿碱合成酶基因已在烟草毛状根基因组中整合并得到表达。烟草毛状根多倍体诱导的最适条件为0.1%的秋水仙素溶液处理36 h,其多倍体诱导率为64.71%。经秋水仙素加倍的烟草毛状根多倍体植株再生的最适宜培养基为MS+6-BA 2.0 mg/L+NAA0.2 mg/L。与对照(二倍体非转化植株)相比,烟草二倍体毛状根再生植株的顶端优势减弱,腋芽增多,叶片变窄;而烟草毛状根多倍体再生植株茎更粗,节间变短,叶色更深,叶片的宽度和厚度均较对照明显增大。根尖细胞染色体压片观察证实,所获得的烟草毛状根多倍体再生植株为四倍体,其根尖细胞染色体数约为4n=96。盆栽实验表明,烟草二倍体毛状根植株和多倍体毛状根再生植株比对照植株延迟约21 d开花。GC-MS检测表明,烟草毛状根多倍体再生植株的烟碱含量比对照及二倍体毛状根再生植株显著提高,分别约为对照及二倍体毛状根再生植株的6.90倍和4.57倍。     

马铃薯StCIPK11的克隆及响应干旱胁迫分析

明确马铃薯StCIPK11在响应干旱胁迫信号传导中的功能和作用机制，为深入研究StCIPK11响应马铃薯抗旱调控的分子机制提供理论依据。利用同源重组法和人工microRNA技术构建马铃薯StCIPK11过表达载体和干扰表达载体，通过根癌农杆菌介导法分别将其转入马铃薯栽培品种‘大西洋’中。RT-qPCR结果表明，过表达植株StCIPK11的表达量是非转基因植株（NT）的11.59和21.76倍，干扰表达植株StCIPK11干扰程度达到78%。经PEG模拟干旱胁迫，过表达植株叶片中丙二醛含量显著高于NT植株，脯氨酸含量、抗氧化酶（超氧化物歧化酶、过氧化物酶）活性均低于NT植株；StCIPK11干扰表达植株则表现出相反的趋势。StCIPK11参与了干旱胁迫应答过程，StCIPK11干扰表达可以降低马铃薯植株对水分胁迫的敏感性。     

聚乙二醇（PEG）浸种对茎瘤芥老化种子活力及其幼苗生长的影响

茎瘤芥品种‘永安小叶’老化种子经聚乙二醇（PEG）浸种后,其种子活力和幼苗生长与PEG浓度有一定的相关性,不同浓度PEG浸种后的茎瘤芥种子膜透性均降低,种子发芽率、发芽势、发芽指数和活力指数增大,幼苗根干重、茎干重和根长也均增大,幼苗叶片中丙二醛（MDA）含量降低。幼苗生长与种子活力的变化趋势一致,PEG浓度小于30%时呈增大趋势,大于30%时呈减小趋势,但均大于不做PEG处理的;幼苗叶片中MDA含量与种子膜透性变化趋势一致,PEG浓度小于25%时,呈降低趋势,大于25%时则呈升高趋势,但这些指标均低于不做PEG处理的。     

多效唑对金钗石斛试管苗生长的影响

多效唑是一种高效的植物生长延缓剂,通过不同浓度的多效唑对金钗石斛试管苗生长的影响进行研究。结果表明多效唑能明显提高金钗石斛试管苗再生芽分化的数量,促进再生苗根的分化,提高生根的数量;同时,多效唑能使再生苗的叶片变短,对再生植株有显著的矮化作用,使植株的茎段变粗,抑制根的伸长生长。多效唑对金钗石斛试管苗生长的影响具有明显的规律性,其作用效果均随着其使用浓度的升高而增强,并在3~4mg/L的范围处达到最强效果,再提高浓度则趋于饱和。     

Nutritional and root development factors affecting growth of tissue culture plantlets of loblolly pine

Tissue culture plantlets of loblolly pine ( Pinus taeda L.) were compared to seedlings to quantify growth and developmental differences. The two plant types were grown in containers in a greenhouse and sampled periodically for twenty weeks. Dry weights and nitrogen and phosphorus concentrations of the shoots and roots were determined every two weeks.
During the twenty weeks in the greenhouse, seedlings grew to a greater size than the plantlets, but the relative rates of growth were approximately equal. Plantlets had significantly lower concentrations of nitrogen and phosphorus per g of shoot dry weight. Seedlings were much more efficient at nutrient uptake per g of dry weight of root. Plantlets had thick, unbranched roots, which were inefficient at nitrogen and phosphorus uptake. Nutrient uptake based on an index of root surface area was equal in the plantlets and seedlings.
The main differences between plantlets and seedlings apparently were related to root system morphology rather than physiological processes. The uptake of nutrients showed the greatest difference between the plant types.     

Polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine (Pinus virginiana Mill.) plantlets

Polyamines have been demonstrated to play an important role in adventitious root formation and development in plants. Here, we present a detailed analysis of influence of exogenously added polyamines on adventitious root development and its relationship to cold tolerance in Virginia pine (Pinus virginia Mill.). Our results demonstrated that polyamines putrescine (Put), spermidine (Spd), and spermine (Spm) at 0.001 mM improve rooting frequency and promote root elongation. Put, Spd, and Spm at 0.01–1 mM decrease rooting frequency and reduce root elongation root elongation. Measurements of diamine oxidase (DAO, EC 1.4.3.6) and polyamine oxidase (PAO, EC 1.4.3.4) activities showed that higher DAO and PAO enzyme activities were obtained when high concentrations of polyamines were applied and when plantlets were treated for 5–7 week at 4°C and 16°C. Survival rate of plantlets increased with the treatment of polyamines at low temperature. Polyamines increased mitotic index of cells in root tips of regenerated plantlet cultured on medium containing 0.001 μM Put, Spd, or Spm, but did not increase mitotic index in tissues of needle tips of the same plantlets. These results demonstrated that polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine plantlets.     

A tripartite culture system for endomycorrhizal inoculation of micropropagated strawberry plantlets in vitro

The objective of the current investigation was to develop a reliable method to obtain vesicular arbuscular mycorrhizae (VAM) in micropropagated plantlets and to determine their influence on growth. An in vitro system for culturing the VA mycorrhizal fungus Glomus intraradices with Ri T-DNA-transformed carrot roots or nontransformed tomato roots was used in this study as a potential active source of inoculum for the colonization of micropropagated plantlets. After root induction, micropropagated plantlets grown on cellulose plugs (sorbarod) were placed in contact with the primary mycorrhizae in growth chambers enriched with 5000 ppm CO₂ and fed with a minimal medium. After 20 days of tripartite culture, all plantlets placed in contact with the primary symbiosis were colonized by the VAM fungus. As inoculum source, 30-day-old VA mycorrhizal transformed carrot roots had a substantially higher infection potential than 5-, 10-or 20-day-old VAM. Colonized plantlets had more extensive root systems and better shoot growth than control plants. The VAM symbiosis reduced the plantlet osmotic potential. This response may be a useful pre-adaptation for plantlets during transfer to the acclimatization stage.     

PP333对离体苹果苗的生长及其生理的影响

在生根培养基中加入0.5或2.0PPm PP333, 明显地减少了苹果离体新梢的鲜重和干重、叶片鲜重和叶面积,增加了比叶重、根的鲜重和干重及根梢鲜重和干重比,并促进了根的形成。PP333对单位叶重量的叶绿素含量没有影响,而增加了单位叶面积的叶绿素含量。经PP333处理后,叶片中的淀粉含量、过氧化物酶活性、蛋白质和游离氨基酸的含量均明显高于对照,可溶性糖与对照差异不明显。     

樟叶越桔组培苗生根和移栽技术研究

为解决樟叶越桔(Vaccinium dunalianum)组培苗生根质量不佳、移栽成活率低的问题,该研究以樟叶越桔继代苗为材料,采用单因子试验从激素类型及浓度、培养基类型和蔗糖质量浓度对其生根的适宜条件进行筛选,进一步研究了不同基质配比对樟叶越桔移栽苗存活率的影响。结果表明:激素类型和浓度、培养基类型对樟叶越桔生根率的影响最大,其次为蔗糖质量浓度;最适合樟叶越桔生根的激素及浓度为IBA2.0 mg·L~(-1)、基本培养基类型为1/4MS、蔗糖质量浓度为15 g·L~(-1),樟叶越桔组培苗最佳生根培养基为1/4MS+IBA 2.0 mg·L~(-1)+活性炭0.1 g·L~(-1)+蔗糖15 g·L~(-1),生根率达100%,平均生根数为每株7.67条;根系呈辐射状、基部无愈伤组织,组培苗生长健壮、叶色浓绿;樟叶越桔组培苗移栽时以全腐殖土基质为佳,成活率为83.7%,植株叶片舒展,生长状况良好。该研究建立的优化体系有效地提高了樟叶越桔组培生根苗的生根率和生根质量,解决了后期移栽成活困难的问题,为优良的樟叶越桔植株规模化生产提供了科学依据和技术支持。     

Transformation of Populus tomentosa by Agrobacterium and Regeneration of Transformed Plantlets

The establishment of efficient transformation system of Populus tomentosa by Agrobacterium is reported. The strains of Agrobacterium used in experiments were: 1. A. rhizogenes R1000, which harboured the Ri plasmid pRiA4b. 2. A. rhizogenes R1000 (pTVK85), which carried the plasmids pRiA4b and pTVK85 Containing supervirulent region. 3. A. tumefaciens C58C1 (pBZ693), the plasmid pBZ693 containing genes 1 and 2. After being cocultured with the bacteria on media containing 0.5 ppm kinetin for 2 days, explants of P. tomentosa were transferred to MS medium containing 500 ppm cefotaxime. Roots appeared on the explants in a week. The roots induced by A. tume[aciens were morphologically different from those induced by A. rhizogenes. The frequency of the explants transformed by A. rhizogenes R1000 (pTVK85) was nearly up to 60%. Some Ri plasmid transformed roots could spontaneously produce adventitious shoots or calli. By adding appropriate plant growth regulators in the media, we could have all of the root lines transformed produce adventitious shoots which would develop into intact plantlets on a hormone-free medium. Some phenotypical differences were observed among clones of the transformed plantlets. Some clones had short internodes, large number of leaves, reduced apical dominance, rich root systems with a great quantity of branches and root hairs, whereas in other clones aboveground parts of plantlets were morphologically normal and only their root systems were different from those of untransformed plantlets. None of the plantlets transformed by A. rhizogenes had the phenomenon of wrinkle leaves and shapes these leaves were analogous to normal plantlets. It was often observed that roots were regenerated from stems above the medium surfaces. Southern analysis on three clones of the putative transformed plantlets by A. rhizogenes R1000 (pTVKS5) showed that two of them were hybridized positively with the probe covering the TL-DNA region of the plasmid pRiA4b.     

Comparative greenhouse study of Eucalyptus grandis in vitro plantlets and half-sib seedlings,II. Dry matter accumulation and relative distribution

In vitro directly micropropagated plantlets from three selected five-year-old Eucalyptus grandis Hill ex. Maiden hybrids were compared to their related half-sib seedlings for growth and growth pattern parameters under greenhouse conditions used for operational seedling production. The oven dry weights were determined from stem, leaf, and root samples collected every 40 days for four times. Relative growth rate, net assimilation rates and shoot:root ratio were calculated. Survival was 98% and 95% for plantlets and seedlings, respectively. Significant differences were observed between parents in terms of shoot and root dry weights and their ratios with similar ranking among plantlets and seedlings, suggesting genetic control over these traits. Plantlets started with significantly higher root: shoot ratios and stem, leaf, root, and total dry weight. Although seedlings had higher relative growth and net assimilation rates, all the initial differences decreased sharply over time.     

牡丹不定根形成相关基因PsARRO-1的克隆及表达分析

不定根发生相关加氧酶基因（adventitious rooting related oxygenase,ARRO—1）被认为是木本植物不定根形成的分子标记之一,属不定根发生起始阶段的特异表达基因。本实验以牡丹‘乌龙捧盛’为材料,运用RT-PCR和RACE相结合的方法克隆得到一个ARRO-1的全长cDNA序列,命名为PsARRO-J（GenBank登录号KJ620008）。PsARRO-1cDNA序列的开放阅读框长度为900bp,编码299个氨基酸。氨基酸序列同源性分析结果显示与拟南芥、毛果杨、苹果、梅花等植物有较高的相似性。利用实时荧光定量PCR对该基因在牡丹‘凤丹白’试管苗及实生苗的表达情况分析表明,PsARRO-1在试管苗及实生苗的根、茎、叶、花中均有不同程度的表达,根中的平均表达量大于其他部位,且实生苗中的整体表达量一般高于试管苗中的表达量。试管苗中,PsARRO-1在根中的表达量变化趋势明显,生根诱导初期表达量平稳且微弱,第10天开始上调表达,第15天和第40天分别出现两次峰值;实生苗中,PsARRO-1在根中的表达量在取样初期就开始快速上升,第10天达到峰值,之后迅速回落。这与牡丹不定根的发生过程基本一致,说明PsARRO-1与牡丹不定根的形成密切相关。     

Controlled mycorrhizal initiation as a means to improve root development in somatic embryo plantlets of hybrid larch (Larix × eurolepis)

This study represents the first report of in vitro mycorrhization of somatic embryo-derived plantlets. Effects of fungi on the rooting of plantlets as well as on their ultra-structure were studied. Embryo-derived plantlets of a hybrid larch ( Larix × eurolepis Henry) were grown aseptically in the presence of four ectomycorrhizal fungi. One month after inoculation with Laccaria laccata, Hebeloma cylindrosporum and Pisolithus tinctorius , the number of plantlets which had developed a well-growing root system was significantly improved compared with controls. Furthermore, root branching was strongly stimulated by L. laccata and H. cylindrosporum , which were more efficient species than P. tinctorius and Suillus grevillei . Development of the root system was concomitantly accompanied by an enhancement of shoot growth, except for plantlets inoculated with S. grevillei , which died after three to six months. At the ultrastructural level, cross-sections of lateral roots made six months after inoculation, showed the presence of a Hartig net in the case of L. laccata and H. cylindrosporum , whereas no such net was observed with the two other fungi. With S. grevillei , some perforating hyphae penetrated the cortical cells and resulted in cell death. Evidence of a relationship between structure and efficiency of the association was found.