萘乙酸与多效唑对茉莉成花及新梢内源激素含量的影响

在茉莉开花前期分别使用不同浓度NAA与PP333均匀喷施于植株茎、叶片等生长部位,对其新梢与花蕾生长及其4种内源激素--生长素(IAA)、赤霉素(GA)、脱落酸(ABA)、玉米素核苷(ZR)的含量变化进行分析.结果表明:(1)NAA处理使茉莉新梢徒长,成花比对照推迟2～4 d;PP333处理的茉莉新梢矮化,而成花比对照提早2～4 d.(2)NAA处理后,茉莉新梢中IAA与ABA含量处理初期较高,后快速下降,后期稳定在较高水平;GA含量稍低于对照,ZR含量降低并稍低于对照.PP333处理后,茉莉新梢中IAA与ABA含量初期较高,而后缓慢下降;GA含量与对照一样快速上升;ZR含量在初期含量较高,后缓慢下降,但较对照稳定在较高水平.(3)PP333处理的茉莉植株新梢中ABA/IAA、GA/IAA、ZR/IAA比值在处理后迅速上升,特别是GA/IAA、ZR/IAA比值明显高于对照,而相应NAA处理的3个比值与对照无明显差异.可见,经NAA与PP333处理能明显调节茉莉新梢及花蕾生长进程,保持较高水平的内源激素GA、ZR、GA/IAA、ZR/IAA比值在茉莉新梢及花蕾生长过程中起关键作用.     

IBA和NAA对山杜英组培苗生根过程中内源IAA、ABA含量变化的影响

本文研究山杜英组培苗生根过程中内源IAA、ABA含量变化规律。结果表明,培养基添加IBA和NAA后,在生根过程中内源IAA、ABA含量变化类似,根点出现前内源IAA、ABA含量一直上升,根点出现后含量开始下降,产生愈伤组织时两种处理的IAA/ABA分别是2.526和3.226。在不添加外源生长素情况下,内源IAA含量一直维持在较低水平,而内源ABA含量一直呈现上升趋势,IAA/ABA始终都在1.211以下。     

油茶未成熟子叶幼胚离体培养成苗的研究

MS培养基适用于油茶未成熟子叶与幼胚的离体培养。在附加2ppm NAA+4PPm BA的培养基上诱导愈伤组织,转入减少生长素或无生长素,提高细胞分裂素的培养基中,可产生大量丛生小苗,每管可达40株以上。也可以在再分化培养基上直接诱导丛生苗。丛生小苗分株培养后,用液体培养基在滤纸桥上诱根,并用透明薄膜室移栽成活。试验表明生长素NAA、IAA对愈伤组织的诱导与分化都有良好效果,BA、ZT与NAA的适当配比是诱导丛生小苗的重要因素。只有绿色或黄绿色的愈伤组织才能分化。培养基含2.4-D,蔗糖5%以上都不利于愈伤组织绿化,也不利于分化。子叶基部与幼胚的培养条件基本相同,而且其分化能力与发育阶段有密切关系,取六月以后,油茶胚乳已完全消失的子叶与幼胚作外植体,较易诱导成苗。除了丛生苗的诱导外,幼胚在怀特培养基附加低浓度的生长素与细胞分裂素上可获得根茎完整的胚苗。但移栽的成活率却低于上述诱根的丛生苗。     

枳多芽苗的诱导

枳幼苗茎段的腋芽在含BA(0.5毫克/升)和NAA(2毫克/升)的MS培养基上能诱导出大量丛生芽,丛生芽苗茎段的腋芽在含BA(0.5-2毫克/升)的MS继代培养基上又可增殖,将无根幼苗分别转至含IAA、IBA和NAA(均为1毫克/升)的MS培养基上则生根形成完整植株。     

植物激素对人参毛状根生长和皂甙含量的影响

就植物激素IAA、IBA、NAA、2,4-D对人参（Panax ginseng C.A.Meyer）毛状根生长及皂甙含量的影响进行了研究。结果表明,4种生长素在适宜的浓度下均可不同程度地促进人参毛状根的生长以及皂甙的积累,同时能影响单体皂甙的分布。NAA和IBA能显著促进毛状根的生长,其中0.500mg/L IBA能显著促进毛状根生长和总皂甙的积累。细胞分裂素6-BA在较低浓度时虽然对生长无明显的促进作用,但对皂甙积累有利,同时显著促进单体皂甙Rb1的积累,增大Rb1在总甙中所占的比例。     

绞股蓝的快速繁殖研究

本文报道以广西四个县所产绞股蓝,在MS基本培养基中进行茎段培养,研究植物激素对器官形成的影响。试验结果表明,细胞分裂素BA明显促进芽的形成和增殖,BA和NAA或GA配合使用有助于苗的形成和生长,不同产地的绞股蓝类型均能诱导形成丛生芽,但分化芽数有差异。诱导生根用1/2 MS附加NAA或IBA,试管苗一年四季移栽均获得较高的成活率。幼苗移栽大田生长良好,当年可获得较好产量。     

欧李“大磨盘”丛生芽的诱导及生根技术

以欧李"大磨盘"的茎尖为外植体,1/2MS和MS为基本培养基,研究了不同生长激素对启动培养的影响,不同培养基对丛生芽增殖的影响,初步建立了欧李"大磨盘"丛生芽的诱导及生根技术。适宜"大磨盘"的启动培养的生长素为IBA,其最佳丛生芽的培养基为MS+6-BA1.0mg/L+NAA0.01mg/L+IBA0.2mg/L,最佳分化培养基是MS+6-BA0.5mg/L+NAA0.01mg/L+IBA0.2mg/L,最佳生根培养基是1/2MS+IBA0.5mg/L+NAA0.1mg/L。     

三倍体毛白杨组织脱分化培养与植株体再生

采用毛白杨三倍体幼叶作为外植体进行组织培养,以MS为基本培养基获得了再生植株。从NAA和IAA与6-BA间进行的18个正交试验中,选择出适宜脱分化培养基MS+6-BA 0.5mg/L+NAA 0.1mg/L,对三倍体毛白杨愈伤组织诱导率为87.6%。5种生长素与6-BA配比的再分化培养基中,12MS+IAA 0.1mg/L+6-BA 0.1mg/L对不定芽的分化诱导率可达到68.5%;MS+6-BA 0.5mg/L+NAA 0.01mg/L的培养基可使单芽直接增殖出7.4个芽。在MS+IBA 1.0mg/L的生根培养基上,试管苗的生根率可达85.7%。     

贯叶金丝桃组织培养的研究

分别以甘肃天水贯叶金丝桃的幼根、幼茎、幼叶为外植体．在1／2MS培养基上附加各类激素,进行贯叶金丝桃的组培实验。研究发现各外植体的增殖速率由高到低分别为幼茎、幼根、幼叶,且得到贯叶金丝桃组培各阶段的最佳培养基成分。诱导愈伤组织的培养基为1／2MS 1．3～1．6mg／L BA 0．2mg／L NAA;培养基1／2MS 1．3～1．6mg／L BA 0．15mg／L NAA有利于不定芽的形成;诱导不定根的培养基为l／2MS IBA0．5～O．8mg／L 蔗糖2．0％。向1／2MS培养基中添加不同的生长素(IAA,IBA,NAA,2．4-D)．在不同浓度梯度的培养基上进行诱导贯叶金丝桃的愈伤组织及不定根的试验,结果表明：生长素IAA,IBA既可诱导愈伤组织,又可以诱导不定根的产生。生长素NAA,2,4-D可诱导产生愈伤组织,但对不定根的诱导作用较差。     

魔芋丛生芽诱导成苗及生根技术研究

采用不同激素配方的1-9号培养基,探索了由魔芋丛生芽诱导试管苗的最佳培养基配方。试验结果表明:MS+6BA1.5 mg.L-1+NAA0.5 mg.L-1+IBA1.0 mg.L-1和MS+NAA1.0 mg.L-1+PP3331.0 mg.L-1两种培养基对于诱导出苗及生根效果最好,添加PP333不仅可加快出苗生根,还可促使试管苗矮壮。     

Auxin influences symptom expression and phytoplasma colonisation in periwinkle infected with periwinkle leaf yellowing phytoplasma

Auxin imbalance was suggested as a key factor in phytoplasma symptom development. Furthermore, remission of the symptoms of phytoplasma‐infected shoots can be promoted by culturing them in vitro in high‐auxin‐containing media. Therefore, effect of spraying 1‐naphthaleneacetic acid (NAA) on infected periwinkle (Catharanthus roseus) with periwinkle leaf yellowing (PLY) phytoplasma was examined. 1‐Naphthaleneacetic acid stimulated symptom development in phytoplasma‐inoculated shoots. Accelerated symptom development was associated with early accumulation of phytoplasmas. Two PATHOGENESIS‐RELATED (PR) genes, CrPR1a and CrPR1b, were induced by PLY phytoplasma infection, and the induction was suppressed by NAA. Therefore, the accelerated symptom development may be due to the suppression effect of NAA on PR‐related defence. However, while NAA promoted symptom development on shoots inoculated with phytoplasma, more non‐symptomatic shoots containing no phytoplasma were observed, suggesting that NAA prevents phytoplasma colonisation in non‐symptomatic shoots. The expression of two genes encoding jasmonic acid (JA) biosynthesis key enzymes, lipoxygenase and allene oxide cyclase, was downregulated in non‐symptomatic shoots of infected plants, and remained downregulated after auxin treatment. Therefore, the auxin‐promoted resistance should be JA independent. Because auxin may promote symptom development of PLY phytoplasma‐infected periwinkles, it may not link to plant resistance to phytoplasma infection.     

Auxin‐treatment induces recovery of phytoplasma‐infected periwinkle

Aims: To test the effect of auxin‐treatment on plant pathogenic phytoplasmas and phytoplasma‐infected host. Methods and Results: In vitro grown periwinkle shoots infected with different ‘Candidatus Phytoplasma’ species were treated with indole‐3‐acetic acid (IAA) or indole‐3‐butyric acid (IBA). Both auxins induced recovery of phytoplasma‐infected periwinkle shoots, but IBA was more effective. The time period and concentration of the auxin needed to induce recovery was dependent on the ‘Candidatus Phytoplasma’ species and the type of auxin. Two ‘Candidatus Phytoplasma’ species, ‘Ca. P. pruni’ (strain KVI, clover phyllody from Italy) and ‘Ca. P. asteris’ (strain HYDB, hydrangea phyllody), were susceptible to auxin‐treatment and undetected by nested PCR or detected only in the second nested PCR in the host tissue. ‘Ca. P. solani’ (strain SA‐I, grapevine yellows) persisted in the host tissue despite the obvious recovery of the host plant and was always detected in the direct PCR. Conclusions: Both auxins induced recovery of phytoplasma‐infected plants and affected tested ‘Candidatus Phytoplasma’ species in the same manner, implying that the mechanism involved in phytoplasma elimination/survival is common to both, IAA and IBA. Significance and Impact of the Study: The results imply that in the case of some ‘Candidatus Phytoplasma’ species, IBA‐treatment could be used to eliminate phytoplasmas from in vitro grown Catharanthus roseus shoots.     

Occurrence and Characterization of a 16SrII‐D Subgroup Phytoplasma Associated with Parsley Witches’ Broom Disease in Iran

During 2010–2013 surveys for the presence of phytoplasma diseases in Yazd province (Iran), a parsley witches’ broom (PrWB) disease was observed. Characteristic symptoms were excessive development of short spindly shoots from crown buds, little leaf, yellowing, witches’ broom, stunting, flower virescence and phyllody. The disease causative agent was dodder transmitted from symptomatic parsley to periwinkle and from periwinkle to periwinkle by grafting inducing phytoplasma‐type symptoms. Expected length DNA fragments of nearly 1800 and 1250 bp were, respectively, amplified from naturally infected parsley and experimentally inoculated periwinkle plants in direct polymerase chain reaction (PCR) using phytoplasma primer pair P1/P7 or nested PCR using the same primer pair followed by R16F2n/R16R2 primers. Restriction fragment length polymorphism and phylogenetic analyses of 16S rRNA gene sequences showed that the phytoplasma associated with PrWB disease in Yazd province belong to 16SrII‐D phytoplasma subgroup. This is the first report of association of a 16SrII‐related phytoplasma with PrWB disease in Iran.     

Molecular Characterization of a Phytoplasma Associated with Potato Witches’‐broom Disease in Iran

Potato plants showing symptoms suggestive of potato witches’‐broom disease including witches’‐broom, little leaf, stunting, yellowing and swollen shoots formation in tubers were observed in the central Iran. For phytoplasma detection, Polymerase Chain Reaction (PCR) and nested PCR assays were performed using phytoplasma universal primer pair P1/P7, followed by primer pair R16F2n/R16R2. Random fragment length polymorphism analysis of potato phytoplasma isolates collected from different production areas using the CfoI restriction enzyme indicated that potato witches’‐broom phytoplasma isolate (PoWB) is genetically different from phytoplasmas associated with potato purple top disease in Iran. Sequence analysis of the partial 16S rRNA gene amplified by nested PCR indicated that ‘Candidatus Phytoplasma trifolii’ is associated with potato witches’‐broom disease in Iran. This is the first report of potato witches’‐broom disease in Iran.     

Witches'Broom of Sarothamnus scoparius: A New Disease Associated with a Phytoplasma Related to the Spartium Witches'Broom Agent

In southern and central Italy, a witches’broom and decline disease of Sarothamnus scoparius has been observed. In affected plants, phytoplasmas were detected by PCR amplification of ribosomal DNA. Restriction fragment length polymorphism analysis of PCR-amplified DNA revealed that the diseased plants were infected by a phytoplasma that is closely related to the spartium witches’broom phytoplasma, a member of the apple proliferation group.     

Auxin stability and accumulation during in vitro shoot morphogenesis influences subsequent root induction and development in <Emphasis Type="Italic">Eucalyptus grandis</Emphasis>

Recent results showed that after 16 months in the field, micropropagated eucalyptus plants have an inferior root system to cuttings. Such differences may be due to the plant growth regulators supplied during the culture stages of standard protocols, which are targeted at optimising plantlet yields and not root quality. This study investigated such a proposal, focusing on auxins in an easy-to-root clone. Initial results showed that the auxin provided in the standard protocol (NAA for multiplication and IBA for elongation) enabled 100% rooting in auxin-free medium, where rooting was faster than on IBA-rooting media. When auxin supply was omitted from multiplication and restricted to NAA or IAA during elongation, rooting in an auxin-free medium was reduced to 68 and 31%, respectively, reflecting the stabilities of these auxins in plant tissues. Additionally, 15% of shoots from the NAA-medium and 65% from the IAA-medium produced roots with altered graviperception. GC–MS analysis of these shoots revealed a relationship between free IAA-availability and altered graviperception. This was further tested by adding the IAA-specific transport inhibitor 2,3,5-triiodobenzoic acid to rooting media with IBA, IAA or NAA, which resulted in 100, 70.9 and 20.6% rooting, respectively. At least 40% of the sampled root tips had atypical starch grain deposition and abnormal graviperception. It is proposed that, at least in this clone, while IBA and NAA can be used for in vitro root induction, IAA is necessary for development of graviresponse.     

Association of a Phytoplasma with Pistachio Witches’ Broom Disease in Iran

Pistachio is an important crop in Iran, which is a major producer and exporter of pistachio nuts. The occurrence of a new disease of pistachio trees, characterized by the development of severe witches’ broom, stunted growth and leaf rosetting, was observed in Ghazvin Province. A phytoplasma was detected in infected trees by polymerase chain reaction (PCR) amplification of rRNA operon sequences. Nested PCR with primer pairs P1/P7 and R16F2n/R16R2 was used for specific detection of the phytoplasma in infected trees. To determine its taxonomy, the random fragment length polymorphism (RFLP) pattern and sequence analysis of the amplified rRNA gene were studied. Sequencing of the amplified products of the phytoplasma 16S rRNA gene indicated that pistachio witches’ broom (PWB) phytoplasma is in a separate 16S rRNA group of phytoplasmas (with sequence homology 97% in Blast search). The unique properties of the DNA of the PWB phytoplasma indicate that it is a representative of a new taxon.     

Molecular detection of Candidatus Phytoplasma spp. causing witches’ broom disease of acid lime (Citrus aurantifolia) in India

Phytoplasma infected acid lime plants in India develop characteristic symptoms like small chlorotic leaves, multiple sprouting and shortened internodes. Leaves drop prematurely and infected branches have distorted twigs resembling witches’ broom appearance which eventually show die-back symptoms. During its first report in 1999, witches’ broom disease identification was made on the basis of symptomatology and electron microscopy. However, molecular techniques have proved to be more accurate and reliable for phytoplasma detection than the conventional methods. During survey in the year 2010 six samples were collected from infected acid lime plants showing typical field symptoms from Vidarbha region of Maharastra. Initially, phytoplasma bodies were observed in phloem tissues of all six symptomatic samples under JEM 100S transmission electron microscope and all these six samples were subsequently screened using different set of phytoplasma specific universal primers by nested PCR, a widely recommended molecular technique for phytoplasma detection. In the present study P1/P7 “universal” phytoplasma-primer set was used for first round of PCR and amplified products were processed separately for nested PCR with three different nested primer pairs viz. R16F2n/R16R2, R16mF2/R16mR1 and fU5/rU3. The presence of phytoplasma was confirmed in all six suspected samples and one representative ~1.2 kb size amplicon was sequenced and deposited in GenBank as Candidatus Phytoplasma species AL-M (JQ808143). This is the first report of PCR based molecular detection of phytoplasma-induced witches’ broom disease of acid lime (WBDL) in India. Further molecular evaluation to determine the identity to the species level is in progress.     

Biochemical and epigenetic changes in phytoplasma‐recovered periwinkle after indole‐3‐butyric acid treatment

Aim: To elucidate the possible mechanism of phytoplasma elimination from periwinkle shoots caused by indole‐3‐butyric acid (IBA) treatment. Methods and Results: It has been shown that a transfer of in vitro‐grown phytoplasma‐infected Catharanthus roseus (periwinkle) plantlets from medium supplemented with 6‐benzylaminopurine (BA) to one supplemented with IBA can induce remission of symptoms and even permanent elimination of ‘Candidatus Phytoplasma asteris’ reference strain HYDB. Endogenous auxin levels and general methylation levels in noninfected periwinkles, periwinkles infected with two ‘Candidatus Phytoplasma’ species and phytoplasma‐recovered periwinkles were measured and compared. After the transfer from cytokinin‐ to auxin‐containing media, healthy shoots maintained their phenotype, methylation levels and hormone concentrations. Phytoplasma infection caused a change in the endogenous indole‐3‐acetic acid to IBA ratio in periwinkle shoots infected with two ‘Candidatus Phytoplasma’ species, but general methylation was significantly changed only in shoots infected with ‘Ca. P. asteris’, which resulted in the only phytoplasma species eliminated from shoots after transfer to IBA‐containing medium. Both phytoplasma infection and treatment with plant growth regulators influenced callose deposition in phloem tissue, concentrations of photosynthetic pigments and soluble proteins, H₂O₂ levels and activities of catalase (CAT) and ascorbate peroxidase (APX). Conclusion: Lower level of host genome methylation in ‘Ca. P. asteris’‐infected periwinkles on medium supplemented with BA was significantly elevated after IBA treatment, while IBA treatment had no effect on cytosine methylation in periwinkles infected with ‘Candidatus Phytoplasma ulmi’ strain EY‐C. Significance and Impact of the Study: Hormone‐dependent recovery is a distinct phenomenon from natural recovery. As opposed to spontaneously recovered plants in which elevated peroxide levels and differential expression of peroxide‐related enzymes were observed, in hormone‐dependent recovery changes in global host genome, methylation coincide with the presence/absence of phytoplasma.     

Detection and Identification of Elm Yellows Group Phytoplasma (16SrV) Associated with Alfalfa Witches’ Broom Disease

In July, 2011, alfalfa plants were observed in Yangling, Shaanxi Province, China with typical witches’ broom symptoms. The presence of phytoplasma was confirmed by transmission electron microscopy and a nested PCR, which amplified a 1.2‐kb fragment using universal primer pairs P1/P6 followed by R16F2n/R2. Sequence, phylogeny and RFLP analyses showed that the alfalfa witches’ broom disease was associated with a phytoplasma of group 16SrV, subgroup V‐B. This is the first record of the 16SrV phytoplasma group infecting alfalfa plants.