蝴蝶兰PhSVP的克隆及其在花发育过程中的表达分析

从蝴蝶兰杂交品种‘大辣椒’中克隆到SVP同源MADS-box基因PhSVP，使用RACE方法获得PhSVP全长，其开放阅读框为687 bp，编码228个氨基酸。蛋白序列多重比对表明PhSVP蛋白包含一个高度保守的MEF2-like MADS结构域和一个中度保守的K-box结构域，且与兰科的SVP/AGL24同源蛋白相似度高，进化树分析也表明PhSVP蛋白与其他兰科植物的SVP/AGL24同源蛋白亲缘关系最近。PhSVP的空间表达模式表明其在营养器官表达量高，在萼片、侧瓣和唇瓣等生殖器官表达量低。进一步对PhSVP在成花转换和花发育过程中花序顶端的表达进行分析，表明PhSVP的转录水平在营养分生组织向花序分生组织过渡的过程中无显著变化，仅在花序分生组织阶段有少量上升，但在花发育早期花分生组织阶段中的表达水平显著升高，而在花发育后期的萼片原基、花瓣原基和合蕊柱原基阶段持续下降。此外还发现A类基因PhAP1在花发育过程中的花序顶端的表达模式与PhSVP相似，而B类基因PhPI和C类基因PhAG在花瓣原基和合蕊柱原基阶段的表达量最高。检测了PhSVP在抽葶期、花蕾期和盛花期的根、叶和花葶中的转录水平，结果表明PhSVP在花蕾期的根中的表达量稍高于其他时期，在不同阶段的叶中的表达水平无显著差异，而在抽葶期花葶中的表达量显著高于花蕾期和盛花期。试验结果暗示蝴蝶兰PhSVP可能起到调控花分生组织状态的维持、避免花器官原基过早分化的作用。     

Reproductive organography of Bougainvillea spectabilis Willd

Bougainvillea spectabilis Willd. is of prime importance for horticulture, as well as potentially for pharmaceutical industries, agriculture and environmental industries. However, its floral development is not yet well understood. A detailed study on floral structure and floral organography in the species was first completed using microscopy of paraffin microtome sections of buds. The results were indicated as follows: first, the three trumped flowers in the cymose inflorescence develop asynchronously. Secondly, Varieties with multi-whorl bracts do not develop any sexual organs, i.e., perianth, pistil and stamens. Thirdly, the wall of the two-loculus anther consists of two kinds of cells: the inner wall, consisting of thick-cytoplasmed cells and the outer wall, consisting of fibrous cells. Fourthly, the pollen grains, with three germination colpi, vary substantially in the form and size in summer under the highest day temperature of 40 °C. Fifthly, the pistil is characterized with betalain-acumulating stylar brush. Followed the developmental course, only one basal ovule is developed in the superior ovary. Finally, organs of one flower develop consecutively from the outer to the inner, i.e., from bracts, to calyx, stamen, and carpel while the three flowers bloomed one by one in one cymose inflorescence. It almost takes 1 week from first bud to the third flower blooming. Our research showed a series of special characteristics of reproduction organography of B. spectabilis which can be useful for understanding its reproduction biology and its sterility.     

Effect of the expression level of an AGAMOUS-like gene on the petaloidy of stamens in the double-flowered lily, ‘Elodie’

Double-flowered lilies, in which stamens are converted to petaloid organs, are valuable for horticulturists. ‘Elodie’ is a double-flowered lily cultivar in which stamens are homeotically converted into petaloid organs in whorl 3. The ‘Elodie’ cultivar shows individual variation in stamen structure and it was therefore classified into the following three types according to the strength of petaloidy of the stamens: weak (type-I), intermediate (type-II), and strong (type-III) phenotypes. The AGAMOUS (AG) gene is a class C floral identity gene in Arabidopsis thaliana that is involved in the formation of stamens and carpels. An AG-like gene was isolated from ‘Elodie’ (LelAG1) and its expression was compared between flower types. The LelAG1 gene was expressed in whorls 3 and 4, but not in whorls 1 and 2 in all flower types. In type-I flowers, LelAG1 was expressed strongly in whorls 3 and 4, while its expression was significantly decreased in whorl 3 in type-III flowers. In type-II intermediate phenotype flowers, the expression level of LelAG1 in whorl 3 was reduced by 60%. These results suggest that the expression level of AG-like genes is correlated with the degree of petaloidy of the stamens.     

Florogenesis of the Mediterranean geophyte Narcissus tazetta and temperature requirements for flower initiation and differentiation

Plant florogenesis is determined by the interaction between the genetics of the individual plant and environmental factors. Flower initiation and development of many studied plant species require low temperatures. However, some geophytes form flowers within the bulb during the summer quiescence period and do not require low temperatures for florogenesis. Narcissus tazetta was employed to study, in detail, the stages of flower development during the summer period, with special emphasis on the effect of temperature on inflorescence initiation. The sequence of morphological processes occurring during floral initiation and development was observed by SEM. During 3 years, the bulbs were subjected to different storage temperatures during the summer, and records were taken of the percentage of apical meristems that shifted from vegetative to reproductive development. Flowering and inflorescence quality were recorded after planting. It was concluded that, under natural conditions, the temperature increase in June, during the growth of the mother plant, leads to the transition of the apical meristem to the reproductive stage. Floral initiation and reproductive development in N. tazetta is promoted by high temperatures with an optimum of 25 °C. Sub- and supra-optimal temperatures (20 and 30 °C) delayed differentiation, while lower temperatures (12 °C) inhibited florogenesis completely.     

6-Benzylamino purine stimulates in vitro shoot organogenesis in Eucalyptus erythronema, E. stricklandii and their interspecific hybrids

In vitro bud and shoot organogenesis was investigated for the ornamental plants Eucalyptus erythronema var. erythronema, E. stricklandii and their interspecific hybrids cv. ‘Urrbrae Gem’ and ‘Hybrid 2.5’ by using 0.0, 0.1, 0.25, 0.5 or 1.0 μM BAP on apex and leaf explants. Callus developed on all explants and increased with all concentrations of BAP without significant differences between BAP concentrations. Buds formed on apex and leaf explants of E. erythronema and E. cv. ‘Urrbrae Gem’ especially with 1.0 μM BAP, but these buds rarely developed into shoots. Bud clusters formed on E. erythronema and E. cv. ‘Urrbrae Gem’ apex and leaf explants whereas E. stricklandii and ‘Hybrid 2.5’ produced fewer, individual buds on the explant. Shoots regenerated from apex explants of all genotypes with all levels of BAP, whereas few shoots of any genotype regenerated from leaf explants regardless of the number of buds formed. Shoots from apex explants could be multiplied successfully. Light microscopy showed meristems developed within the callus, and at the callus and bud surfaces. However, few shoots developed considering the level of bud and meristem formation. This report is the first for successful shoot organogenesis and multiplication in an ornamental eucalypt.     

Changes in morphological phenotypes and essential oil components in lavandin (Lavandula × intermedia Emeric ex Loisel.) transformed with wild-type strains of Agrobacterium rhizogenes

Hairy roots were induced from leaf-derived calli of lavandin (Lavandula × intermedia Emeric ex Loisel.) by infection with wild-type strains of Agrobacterium rhizogenes, A-5 (MAFF 02-10265) and A-13 (MAFF 02-10266). A-5-inoculated calli formed hairy roots more efficiently than A-13 ones. The transgenic shoots could be obtained from hairy root segments mediated by each Agrobacterium strain. However, different plant growth regulators were required for efficient adventitious shoot formation in each strain. In A-5, the most efficient adventitious shoot formation rate of 23.8% was observed in a medium with 4.4 × 10⁻⁶ M of 6-benzylaminopurine. On the other hand, a significantly higher rate of 13.2% was detected in a medium with 4.0 × 10⁻⁷ M of N-(2-chloro-4-pyridyl)-N′-phenylurea in A-13. Most of the regenerated plants showed dwarfism with closed internodes and extensive lateral branching, which were typical characteristics of ‘hairy root syndrome’. On the other hand, only nine of the 45 regenerated plants formed flower buds in early June, a delay of about one month compared with nontransgenic regenerated plants. The floral stalks and spikes of these plants were very short, resulting in a compacted form. Many regenerants showed a significantly lower productivity of essential oil than nontransgenic regenerants. Moreover, the relative percentage of the linalyl-cation-derived compounds, linalool and linalyl acetate, decreased in most of the regenerated plants. Compact plants with the ability of flower bud formation are assumed to be valuable not only for lavandin breeding, but also for clarifying the interaction between rol genes expression and essential oil production.     

Change in flower morphology of Torenia fournieri Lind. induced by forchlorfenuron application

We induced various flower morphologies in torenia (Torenia fournieri Lind.) by the application of forchlorfenuron (CPPU). Those morphologies were the combination of four basic morphological changes, the development of serrate petals, incised petals, a paracorolla, and an increased number of floral organs. These morphological changes occurred systematically depending on the floral stage at the time of CPPU application. Serrate petals were induced when CPPU was applied during the stages of corolla development, whereas application at younger stages induced petal incision. The serrate petal margin resulted from preferential proliferation of cells around the vascular bundles, whereas petal incision likely resulted from the lateral outgrowths of petal. A paracorolla was induced at the adaxial petal face when CPPU was applied between the sepal development stage and early corolla development. The paracorolla appears to have arisen from the lateral outgrowths of the stamen. The numbers of stamens, petals, and sepals increased when CPPU was applied at and before the differentiation of sex organs and the corolla. Enlargement of the floral meristem probably caused this increase. Application of N⁶-benzylaminopurine and zeatin did not induce these morphological changes.     

Effects of lipoxygenase on the corm formation and enlargement in Gladiolus hybridus

To investigate the effects of lipoxygenase (LOX1) on the corm formation and enlargement in Gladiolus hybridus, the field and in vitro researches were conducted to detect the growth and development course, the LOX1 activity, the contents of endogenetic methyl jasmonate (MJ), sucrose, starch and cellulose, as well as LOX isozymes and the expression of its genes in ‘Rose Supreme’ and ‘Advanced Red’. The field investigation showed that the highest activities of LOX1 activity and MJ content in stolons, but not detected in leaves, which were higher in ‘RS’ than in ‘AR’. And the contents of sucrose, starch and cellulose in corm and cormels showed different increase during growth and development of Gladiolus corm. The in vitro investigation showed that the salicylhydroxamic acid (SHAM) treatment caused obviously delayed corm formation, decrease of fresh weight and diameter, and percentage of corm formation; reduced contents of total dissoluble protein, MJ and carbohydrates, and LOX1 activity. Additionally the activity of LOX isozymes became weaker, with two isozyme bands disappeared. In contrast, these physiological and biochemical indexes were increased under MJ treatment. RT-PCR also demonstrated that the expression of LOX1 gene was inhibited under SHAM treatment, induced under MJ treatment. These results suggested that LOX1 might regulate the growth and development of Gladiolus corm through affecting JAs biosynthesis cycles, and then resulting in carbohydrate accumulation. The corm formation could be improved by MJ, but suppressed by SHAM, in which, MJ 0.5 mmol/L was the optimum concentration for promoting corm formation and enlargement in Gladiolus.     

Intergeneric hybridizations between Opisthopappus taihangensis and Chrysanthemum lavandulifolium

For the first time, reciprocal intergeneric hybridizations were produced between Opisthopappus taihangensis and Chrysanthemum lavandulifolium without emasculation. About 20% seed set was similarly obtained from reciprocal hybridization. Only 5 and 6 of the viable plants observed from 45 and 78 seedling survived present some conspicuous intermediate characteristics. Phenotypic evaluation among the progenies of the parents and the putative hybrids was performed carefully since an average of 5.3% seed set was produced in the type of self-pollination using pollen from the same flower and >10% seed set was similarly obtained in the types of self-pollination using pollen from different flowers in a plant and flowers in individual plants from different seeds. One individual of each hybrid shared the inflorescence habit with the pollen plant was confirmed further by the sequence of ncpGS. The two hybrids might be used as bridges of breeding of multi-generic hybrids.     

Vegetative propagation of Alstroemeria in vitro

Young actively growing tissue explants from Alstroemeria inflorescence stem taken at a distance of 1–2 mm below the apex are capable of regenerating buds and roots from which small plantlets can be established. Root and bud regeneration was directly from the stem tissue and not from the callus. White's medium supported growth and regeneration as well as Murashige & Skoog's medium. A higher ratio of auxin to cytokinin resulted in root regeneration, while the reversed ratio promoted bud differentiation. Plantlets were obtained upon bud subculture on a low sucrose medium supplemented with indoleacetic acid but without kinetin.     

Carbohydrate changes in kiwifruit buds during the onset and release from dormancy

Carbohydrates were measured in axillary bud meristems from kiwifruit (Actinidia deliciosa (A.Chev.) C.F. Liang et A.R. Ferguson cultivar Hayward) to determine whether concentrations changed during winter dormancy. Buds were collected from kiwifruit vines growing in four sites, which spanned the climatic range that kiwifruit are grown in New Zealand, and from vines where the time of budbreak had been manipulated using hydrogen cyanamide. During winter, sugars comprised 78% of measured carbohydrates in meristems, with sucrose accounting for more than 86% of sugars detected (up to 200 mg g DW⁻¹). Starch concentrations in bud meristems showed little change during winter. Meristem sucrose and hexose concentrations increased rapidly from autumn until mid-winter, and this was correlated with leaf abscission. Sugar accumulation ceased in mid-winter and concentrations remained stable until spring, suggesting that bud meristem activity was low. Four weeks before visible bud movement, a marked decline in sucrose and increase in hexose concentrations, indicated activity in the meristems had increased prior to budbreak. The patterns of seasonal change and peak sucrose concentrations in meristems were similar at all sites. The only difference was in the timing of events, with changes in meristem sugars occurring earliest in vines growing at the coolest site, coinciding with their earlier cessation of and resumption of vine growth. Advancing budbreak, by applying hydrogen cyanamide to vines in late winter, also advanced changes in sugar concentrations within meristems. This work suggests that the concentration of sucrose in kiwifruit bud meristems has utility as an indicator of whether they are being induced into, maintained or released from winter dormancy.     

The responses of early-flowering chrysanthemums to daylength

In a study of photoperiod response 30 early (summer)-flowering chrysanthemum cultivars from Britain and Japan reacted as quantitative short-day plants. Both flower bud initiation and development occurred more rapidly in short days (SD) as compared with long days (LD). The early-flowering cultivars differed from late-flowering chrysanthemums in that lateral flower buds, and often the terminal flower buds, develop to anthesis in LD. Internodes were shorter in SD and flowers had fewer florets. There was an increased number and proportion of disc florets in SD in all cultivars except one. Two cultivars, ‘Mezame’ and ‘Pennine Yellow’, closely approached day-neutrality.All cultivars produced fewer leaves in LD on upper lateral shoots than on main stems. Either the presence of the terminal flower bud induces earlier flowering of the upper lateral shoots, or the change which causes the terminal apical meristem to initiate a flower in LD influences the axillary meristem also.SD could be used to speed up the flowering of glasshouse-grown crops of early-flowering cultivars. The advantages and disadvantages which may be associated with treatment at different stages of growth are discussed.     

Ethylene influences development and flowering of Ptilotus spp. in vitro and ex vitro

The genus Ptilotus has immense potential for ornamental horticulture but its commercial development has been hindered by propagation limitations. Poor seed quality and germination are reported. Cutting propagation is limited by cutting supply as the juvenile phase of Ptilotus is short. Micropropagation has been used in an attempt to overcome these difficulties but explants become floral in vitro and this causes plantlets to elongate. Ethephon has been used to control flowering of stock plants of many ornamental species. This study investigated the effect of ethephon applied to young (3-week-old, deflasked from tissue culture) and mature (1-year-old) Ptilotus plants in a greenhouse. A system of applying gaseous ethylene at 0, 100, 200 and 300 mg l⁻¹ to the headspace of in vitro plantlets in glass jars was developed and the response of in vitro plantlets to ethylene studied. One-year-old Ptilotus plants were treated with 500 mg l⁻¹ ethephon 2 days before pruning or 1 or 2 weeks after pruning. Ethephon application 2 days before pruning decreased the number of inflorescences and increased the number of shoots (compared to the control) but was phytotoxic. Ethephon applications of 150 or 300 mg l⁻¹ applied weekly or fortnightly to 3-week-old plants deflasked from tissue culture reduced plant height and number of inflorescences and at low concentrations increased the number of new shoots. A fortnightly application at 150 mg l⁻¹ is recommended. Previous reports on the effects of ethylene on inflorescence production on plantlets in vitro are limited. Our study showed that exposure of in vitro plantlets of P. nobilis to ethylene gas at 100 mg l⁻¹ for 1 h significantly increased the number of shoots and plant height but this did not occur for plantlets of P. spicatus. Plantlets of P. spicatus exposed to transient ethylene at 200 and 300 mg l⁻¹ showed significantly greater rooting (52.4%) than the control (13.6%).     

Study of some factors affecting the growth and development of myoga (Zingiber mioga Roscoe)

In order to optimise production of the edible flower buds produced by myoga (Zingiber mioga Roscoe) a greater understanding of the growth and development of the plant and factors influencing flower initiation and development are required. The vegetative growth phase in myoga was characterised by a distinct period of pseudostem development from the planted rhizome piece followed by an extended period of rhizome growth from both the base of pseudostems and the planted rhizome piece. The transition from pseudostem formation to the initiation of rhizome growth occurred at the same time as the dry weight of the planted rhizome piece ceased to decline. Flower bud initiation and development occurred over an extended period, beginning soon after the commencement of new rhizome growth and ending prior to foliage senescence. Flower buds were observed at the terminal meristem of first, second, third and fourth order rhizomes. Increasing temperatures stimulated both increased vegetative growth and flower bud initiation and development. Low flower bud yields recorded under conditions of low temperature were the result of reduced rate of floral initiation and not abortion of flower buds. Increased shading of plants grown under glasshouse conditions resulted in reduced flower bud yield and similarly the response resulted from decreased initiation rather than abortion.