Histone H4 gene expression and morphological changes on shoot apices of strawberry (Fragaria × ananassa Duch.) during floral induction

To investigate flower induction in June-bearing strawberry plants, morphological changes in shoot apices and Histone H4 expression in the central zone during flower initiation were observed. Strawberry plants were placed under flower inducible, short-day conditions (23 °C/17 °C, 10 h day length) for differing number of days (8, 16, 20, 24 or 32 days) and then these plants were transferred to non-inducible, long-day conditions (25 °C/20 °C, 14 h day length). The shoot apices of plants placed under short-day conditions for 8 days were flat, similar to shoot apices of plants in the vegetative phase of development, and Histone H4 was not expressed in the central zone during the experimental period. On the other hand, the shoot apices of plants placed under short-day conditions for 16 days remained flat, similar to shoot apices of plants placed under short-day conditions for 8 days, but Histone H4 was expressed in the central zone at the end of the short-day treatment. Morphological changes in the shoot apices of these plants were observed 8 days after the change in day-length. These plants developed differentiated flower organs after they were grown for another 30 days under long-day conditions. These results indicate that changes in the expression pattern of the Histone H4 gene occur before morphological changes during flower induction and that the expression of the gene in the central zone can be used as one of the indicators of the flowering process in strawberries.     

The effect of sucrose on the expression of the VvTFL1 and VFL genes during flower development in the “Xiangfei” grapevine

VFL and VvTFL1 genes expression patterns and the effects of sucrose on the expression of VFL and VvTFL1 genes in different organs of the “Xiangfei” grapevine (Vitis vinifera L.) were investigated. VvTFL1 gene expression was detected in the meristem of the apical bud and lateral bud, but was not detected during inflorescence differentiation and flower organ development. After sucrose treatment, VvTFL1 gene expression increased in the apical bud, but decreased in the lateral bud. These results suggested that the VvTFL1 gene might be mainly involved in the apical growth process of shoots, and exogenous sucrose had an effect on the VvTFL1 gene by increasing shoot apical meristem initiation of apical buds. The VFL gene was expressed primarily during inflorescence differentiation and early flower organ development, but it gradually reduced in later flower development. After sucrose treatment, VFL gene expression increased in the inflorescence and small or middle flower, but a little change was seen in the large flower. These results suggested that the VFL gene plays important roles in the initiation of inflorescence meristems and the morphological formation of flower organs. Exogenous sucrose had an effect on VFL gene expression at the early stage of flower development.     

Identification of drought-induced genes from the leaves of Rangpur lime (Citrus limon (L) Osbeck)

Water stress is one of the limiting factors for citrus production and Citrus species show great variation in their response to drought stress. Although the majority of Citrus rootstocks are sensitive to water stress, Rangpur lime (Citrus limon (L) Osbeck) (RL) shows a high degree of drought tolerance. Therefore, it has been used as a rootstock in drought-prone environments, but mechanisms of drought tolerance are not yet known. In this study, to explore the mechanisms of drought adaptation and tolerance, a subtractive cDNA library was constructed from the leaves of 14-day drought-stressed and non-stressed RL for identification of drought-induced genes. 285 cDNA sequences were obtained from randomly selected clones from the subtracted library containing the drought-induced genes. The expression analyses of 200 cDNAs in 14-day drought treated and untreated RL by macroarray hybridisation revealed that the expression of 56 cDNAs increased two to 11-fold. 30 non-redundant drought-induced genes were identified from these cDNAs and drought induction of eight selected genes was confirmed by a real-time RT-PCR assay suggesting that expressions of these genes were regulated by drought-stress. Genes identified in this study were mostly related with cell rescue and defense pathways involved in drought adaptation and tolerance of RL.     

Effect of photoperiods before,during and after vernalization on flower initiation and development and its varietal difference in Japanese bunching onion (Allium fistulosum L.)

Summary

To control the bolting of Japanese bunching onion (Allium fistulosum L.) photoperiodically, the effect of photoperiods before, during and after vernalization on flower initiation and development and the varietal differences were investigated using the two mid-season flowering cvs Kincho and Asagi-kujo, and a late-season flowering cv. Cho-etsu. A long-day photoperiod (LD, 16 h) given before vernalization inhibited flower initiation. Especially, the bolting rate of ‘Asagi-kujo’ decreased by about a half, compared with the short-day photoperiod (SD, 8 h). The interaction between the effect of night temperature (3°C, 7°C, 11°C or 15°C) and the effect of the photoperiod (SD and LD) during vernalization was also investigated. In ‘Kincho’, LD did not affect flower initiation at 3°C, but inhibited flower initiation at 7°C, 11°C and 15°C. In ‘Asagi-kujo’, flower initiation was significantly inhibited by LD under all temperature conditions. This inhibitory effect was stronger at 11°C and 15°C than at 3°C and 7°C. In ‘Cho- etsu’, LD significantly inhibited flower initiation at 3°C and 7°C, and flower initiation rarely occurred at 11°C and 15°C. In this study, generally, LD during vernalization inhibited flower initiation in all cultivars. Thus Japanese bunching onion required a short-day photoperiod in flower initiation, which was stronger in ‘Asagi-kujo’ and ‘Cho-etsu’ than in ‘Kincho’. From these results, we conclude that low temperature and a short-day photoperiod complementarily induce flower initiation in Japanese bunching onion. Varietal differences exist in the requirement of low temperature and a short-day photoperiod: the primary requirement in ‘Kincho’ is low temperature and that in ‘Asagi-kujo’ is a short-day. After flower initiation, the early stage of flower development is day-neutral, and after the floret formation stage, a long-day photoperiod promotes flower development and elongation of the seedstalk.     

Short days and temperature effects on growth and flowering in strawberry (Fragaria × ananassa Duch.)

Summary

‘Korona’, ‘Elsanta’, ‘Bounty’ and ‘Senga Sengana’ strawberry (Fragaria × ananassa Duch.) plants, were placed at constant temperatures of 9, 15 or 21°C and daylengths of 8 h (short day) or 24 h (long day). The plants were given different numbers of short-day (SD) cycles, and flowering and growth were studied. ‘Korona’ and ‘Elsanta’ were responsive to both short-day treatment and temperature, with optimum flowering at 15°C and 24 SD. ‘Bounty’ was more responsive to temperature, inducing flowers independently of the number of SD cycles at 9°C and 15°C. In ‘Senga Sengana’ flowering was induced independently of temperature and the number of SD cycles, indicating that it had a stronger dependence on other environmental effects. The effect of the number of short-day cycles and the temperature on vegetative growth variâtes such as the number of stolons and daughter plants, the length of flower trusses and petiole length were also studied.     

Effects of photoperiod on shoot elongation and endogenous gibberellins in the glasshouse carnation

Plants of glasshouse carnation were grown in photoperiodic cycles comprising 8 h natural daylight followed by either 16 h of darkness (short-day treatment) or by 16 h of low-intensity lighting from tungsten filament lamps (continuous-light treatment). When plants were transferred from short days to the continuous-light treatment, rates of shoot elongation were increased and flower initiation was promoted. Shoot tips of plants grown in continuous light yielded greater amounts of diffusible gibberellin-like substances than shoot tips of plants grown in short days. Yields of diffusible gibberellins increased with increases in the duration of the continuous light treatment up to 28 days, by which time flower initials were present. Application of GA₃ to the plants resulted in increased rates of shoot elongation, but no effect on flowering was observed.     

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.     

Identification of differentially expressed genes in fragrant rose Jinyindao with suppressive subtraction hybridization

To investigate the floral fragrance new genes, scent mutant of rose was used here. The suppressive subtraction hybridization (SSH) technique and micoarray analysis of the clones were used to isolate the cDNA fragments, which showed differential expression between the rose scent mutant ‘Wangriqinghuai’ and wild type ‘Jinyindao’ (Rosa × hybrida), and RT-PCR was used to identify up-regulated expressed genes. 16 positive contigs of JSSH were obtained. Some ESTs such as RcOMT1, RcOMT2, RhMYB92 and RhGP1 were known to regulate scent metabolism, and 5 ESTs with no homology in NCBI may represent new genes involved in rose flower fragrance metabolism. SSH technique combined with cDNA micoarray would be useful for analysis and isolation of the genes related to rose floral scent.     

月季突变体抑制差减杂交cDNA文库构建及分析

为了分析月季花色花香突变机理和揭示花色花香代谢的相关基因, 利用抑制性消减杂交技术分离了月季红花无香突变体‘往日情怀’ (以下简称突变体) 与其野生型金黄色浓香品系‘金银岛’ (以下简称野生型) 之间表达差异cDNA片段。分别以突变体作为驱赶子, 野生型为检测子, 以及以野生型作为驱赶子, 突变体为检测子建立了两个差异表达cDNA文库WSSH和JSSH, 分别代表在突变体和野生型中特异表达的cDNA; 再经文库高密度点阵膜的杂交差示筛选分析, 在WSSH库中获得特异表达的27个阳性克隆, 在JSSH文库中得到25个阳性克隆。差异表达克隆测序后经BLAST比对分析发现WSSH文库中含有与红花突变体的花青素积累直接相关的CHS、DFR、细胞色素P450加单氧酶、乙二醛酶Ⅰ、己糖转移因子、MYB1 转录因子、S - 腺苷蛋氨酸转移酶、ADR等花色相关EST; JSSH文库中含有与野生型花香形成相关的月季甲基间苯二酚O - 甲基转移酶、转醛醇酶、Acyl-CoA 结合蛋白、钙调素结合蛋白、MYB92转录因子的EST以及导致芽变的Ty1-cop ia-like逆转座子。     

Molecular characterization of an S-adenosylmethionine decarboxylase gene from floral organ differentiating axillary bud in calamondin (× Citrofortunella mitis)

Summary

Calamondin (× Citrofortunella mitis J. Ingram & H. E. Moore) is known for its ability to develop floral organs to flower in the adult phase during the four seasons. To provide information on molecular events during floral differentiation from axillary buds, we constructed a cDNA library from floral differentiating axillary buds of adult phase calamondin. Ninety-six cDNA clones were randomly selected from this library and sequenced. Ninety-six nonredundant ESTs were identified. Two clones encoded S-adenosylmethione decarboxylase, a key enzymes in polyamine biosynthesis. CmSAMDC 1 (Citrofortunella mitis s-adenosylmethionine decarboxylase 1) contained an 1083 bp ORF that encoded a putative SAMDC precursor of 361 amino acids. Northern blot analysis revealed that CmSAMDC 1 was expressed in axillary buds prior to floral differentiation and in axillary buds immediately after floral differentiation, in immature flower (5 mm), in fully developed flowers (120 mm long), in floral parts (petals, pistils, and stamens) of trees in the adult phase of development, but not in leaves or axillary buds of juvenile phase nucellar seedlings or leaf tissue from trees in adult phase of development. In situ hybridization revealed expression of the CmSAMDC 1 gene in the floral apex of axillary buds after differentiation, and in the phloem of axillary buds and petioles. These results suggest that SAMDC could play a role in actively differentiating and developing reproductive and vegetative organs.     

Isolation and expression analysis of differentially expressed genes in stem tissue of the Greek lemon cv. Adamopoulou

Lemon (Citrus limon (L.) Burm. f.) is susceptible to mal secco, a serious vascular disease caused by the fungus Phoma tracheiphila (Petri) Kant. and Gik., as well as low temperatures. The greek lemon cultivar Adamopoulou, thought to be derived from the Portuguese cultivar Lisbon, exhibits enhanced resistance to mal secco and cold as opposed to cv. Lisbon. Suppression subtractive hybridization (SSH) was employed for the isolation of differentially expressed genes in lemon stem tissue. A subtractive cDNA library was constructed and a total of 296 clones were sequenced. The obtained sequences were edited, resulting in 56 non-redundant ESTs. Sequence analysis revealed homology to previously identified genes involved in defense mechanisms against biotic and abiotic stresses, as well as sequences with no significant similarity in the GenBank. Selected ESTs were analyzed by real-time PCR for confirmation purposes. This analysis revealed significant expression differences between the two cultivars for genes expressing allantoinase, ultraviolet-B-repressible protein, 4-coumarate:CoA ligase and other proteins that are known to be upregulated under biotic and abiotic stress conditions.     

Inhibition of flower stalk elongation and abnormal flower development by short-day treatment in a Japanese variety of Chinese chive (Allium tuberosum Rottler ex Sprengel)

Chinese chive is produced year-round in Japan, therefore it is important to demonstrate the relationships of short-day (SD) conditions with flower stalk elongation and flower formation for optimizing the cropping system, e.g. the increase in harvests per year. To clarify the influence of short photoperiod on flower stalk elongation and flower formation in Chinese chive, 8 h SD treatments were applied at different developmental stages of flower stalk elongation and flower formation. When the SD treatment started from vegetative or floral-initiated stages, the earlier the SD treatment started, the fewer flower stalks appeared. Also, the earlier the SD treatment started, the less the flowers bloomed and the more the flower stalk elongation was inhibited at the end of SD treatments. Many involucres did not open and withered with death of florets in SD when the SD treatment started between the umbel or flower bud differentiation and the perianth to stamen-formation stages. Also, all or part of the florets aborted and there were no complete inflorescences in the later SD treatments. We found that, in Chinese chive, the development of flower stalk elongation and flower formation were inhibited with the earlier SD treatment, after vegetative or floral-initiated stages. Furthermore, it is considered that Chinese chive needs long-day (LD) for the flower stalk elongation and inflorescence formation after the initiation of the flower bud. The plant has a qualitative LD requirement with the same photoperiodic requirement for both flower bud initiation and flower development.     

金钗石斛花芽cDNA表达文库的构建及鉴定

 cDNA文库是分离、筛选基因的重要平台。利用SMARTTM cDNA library construction技术, 成功构建了金钗石斛花芽cDNA表达文库。结果表明该文库的克隆数为3.02 ×10⁵ pfu, 插入片段的大小在0.5～2.5 kb范围, 插入片段的重组率接近100% , 说明所构建的cDNA文库质量较好。该文库的构建为克隆与金钗石斛花芽分化相关功能基因奠定了基础。     

Changes in carbohydrate contents in shoot tips,leaves and roots of strawberry (Fragaria × ananassa Duch.) during flower-bud differentiation

Flowering is an important step in crop production. Under flower-inducing conditions, biochemical or physiological changes can be recognized. Changes in carbohydrates have an important role in flower development in plants; however, carbohydrate changes during flower bud differentiation in strawberry have not been thoroughly investigated. In this study, runner plants potted in 18 cm diameter pots and grown under non-inducing conditions (28 ± 3/22 ± 3 °C day/night; 16 h day length). When the plants were established, half of the plants then were put under inducing conditions (23 ± 3/13 ± 3 °C day/night; 8 h day length). After the induction period of 21 short-day cycles, plants were brought to non-inductive conditions again. In order to evaluate the carbohydrate changes during flower differentiation, shoot tips, leaves and roots were sampled from four replicated plants collected weekly for the period of 7 weeks. Sucrose, glucose and fructose contents were determined by HPLC, and starch by the anthrone method. The results obtained indicated that the most abundant soluble sugar in all organs tested was sucrose. Sucrose in the shoot tips of induced plants at 42, 56 and 70 days after the start of the short-day treatment were significantly higher than corresponding time in non-induced plants. However, the glucose, fructose and starch contents in shoot tips, leaves and roots of non-induced plants in most sampling dates were greater than those of induced plants. In other words, the shoot tips (bud) of induced plants acted as strong ‘utilizing sink’ and preferentially metabolized carbohydrates rather than storing them. It seems that non-structural carbohydrate contents in shoot tips, leaves and roots of strawberry may have an important role in flower-bud differentiation.     

Genome-wide identification and involvement of litchi SPL genes in flowering in response to cold and leaf maturity

Litchi (Litchi chinensis Sonn.) flowers from shoot apical meristems after exposure to prolonged cold differently depending on maturity. The molecular mechanism defining that interaction of cold with maturity is not fully understood. In the present study, potted litchi plants with terminal flushes at mature and turning stage were treated with 60-days of cold. The mature plants, but not the turning plants, were competent to flower, indicating the importance of terminal flush maturity on litchi flowering. This result could be explained by the difference in accumulation of soluble sugars in the apical leaves on terminal flushes. To explore this link, the miR156/SPL (SQUAMOSA-Promoters Binding Protein-like) module, a dominant sugar-mediated flowering regulator, was investigated. Eighteen SPL homologues were identified, 11 of which were up-regulated during the transition from juvenile to adult. Ten LcSPLs were highly expressed in response to cold, but only LcSPL1 and LcSPL2 correlated with the age-dependent flowering in response to cold. Further, 12 LcSPLs, putative targets of LcmiR156, were subjected to transient co-expression assays in tobacco leaves. LcSPL3 and LcSPL10 showed direct binding to the LcFT1 promoter in vitro and in vivo. This trans-activation was verified by yeast one-hybrid (Y1H) assays.