WRKY25过量表达导致拟南芥在长光照下开花提前

WRKY基因家族是主要存在于植物中的一大类转录调控因子,拥有很多家族成员.拟南芥WRKY25属于第Ⅰ类WRKY蛋白,参与植物生物和非生物胁迫反应.通过GUS染色和qRT-PCR发现WRKY25基因主要在根,叶和茎生叶中表达.过量表达WRKY25的转基因植株在长光照下比野生型拟南芥提前开花.通过RT-PCR检测与开花时间相关基因发现,AP1基因的表达量在培养21 d和27d的WRKY25过量表达植株中上调,由此推测WRKY25很可能通过增强AP1的表达来影响开花.     

拟南芥WRKY68转录调控因子的表达谱分析

WRKY基因家族是主要存在于植物中的转录因子,拟南芥中至少有74个成员。根据锌指结构特征和WRKY结构域的数目,可以将WRKY转录因子分为三大类。拟南芥WRKY68属于第Ⅱ类WRKY蛋白。通过GUS染色和qRT PCR分析各组织部位的表达情况,发现WRKY68在根中的表达量是最高的,其次是幼嫩的叶片和老的荚果中。各种处理条件下的表达水平显示,IAA和高温处理后,WRKY68的表达明显上调,PstDC3000、JA、SA、NAA轻微诱导WRKY68的表达,而Botrytis、NaCl、甘露醇、PEG、脱水、ACC、ABA抑制WRKY68的表达,根据以上实验结果,我们推测WRKY68可能参与生长素和温度调控的植物形态建成及发育过程。     

枇杷成花相关基因EjWRKY12的克隆及表达分析（“园艺植物种质资源挖掘与利用”专题约稿）

以库尔勒香梨为试材,在大蕾期喷施不同浓度乙烯利,调查库尔勒香梨萼片脱落率,观测果实生长发育进程和落果情况,测定成熟期果实品质,并基于主成分分析法对乙烯利处理下的果实品质进行综合评价,以揭示乙烯利对库尔勒香梨果实生长发育进程和果实品质的影响,为筛选最优乙烯利浓度应用于提高库尔勒香梨果实品质提供理论依据和技术支持。结果表明：(1)2021年自然生长状态下(对照)库尔勒香梨萼片脱落时间为9 d,不同浓度乙烯利处理后脱落时间会缩短1~2 d,且随着处理浓度梯度的升高,萼片脱落时间和萼片脱落高峰提前。(2)库尔勒香梨果实生长发育期约为130 d,乙烯利处理下和自然状态下的果实生长发育均呈“S”型曲线的变化趋势;乙烯利处理明显加快了果实成熟的速度,果实在花后110 d左右已达到成熟标准,从而可以确定合理的采收时间,并以300 mg·L^-1乙烯利处理下脱萼果和宿萼果的纵横径和单果重增长最大;乙烯利处理下库尔勒香梨在第二次果实膨大期间落果较多,在花后50 d和110 d左右出现落果高峰,并以200 mg·L^-1乙烯利处理对库尔勒香梨果形指数和落果率的影响最明显。(3)300 mg·L^-1乙烯利处理对库尔勒香梨果实品质影响最大,宿萼果纵、横径和单果重较对照分别显著增加5.02%、10.90%和11.40%,脱萼果鲜干比重、可溶性固形物含量、VC含量和可溶性糖含量比对照分别显著提高5.67%、12.03%、21.48%和10.22%,脱萼果硬度和可滴定酸含量比对照分别显著降低10.10%和19.75%。(4)主成分分析显示,各浓度乙烯利处理下库尔勒香梨综合品质得分从高到低依次为300 mg·L^-1脱萼果、300 mg·L^-1宿萼果、200 mg·L^-1宿萼果、200 mg·L^-1脱萼果、100 mg·L^-1脱萼果、100 mg·L^-1宿萼果、CK脱萼果和CK宿萼果。研究发现,大蕾期喷施乙烯利促进了库尔勒香梨萼片的脱落,明显加快了果实的生长发育进程,并可有效提高果实品质,且以300 mg·L^-1乙烯利处理对库尔勒香梨果实品质改善效果最优。     

EFFECTS OF SINGLE AND MULTIPLE EXPOSURES OF FERULIC ACID ON THE VEGETATIVE AND REPRODUCTIVE GROWTH OF PHASEOLUS VULGARIS BBL-290

Phaseolus vulgaris BBL-290 plants were grown in growth chambers in the Southeastern Plant Environment Laboratory and exposed to either single (at seedling, flower, or podfill) or multiple (biweekly or weekly) treatments of ferulic acid (FA). In the first experiment, plants were harvested one week after FA treatment (0, 1.0, 2.0 mM) and at final harvest (56 days old). FA delayed leaf expansion during the seedling and flowering stages. The total plant leaf area and the plant dry weight of plants treated with 1.0 and 2.0 mM FA as seedlings were reduced one week after treatment by 38–48%. The total plant leaf area and the plant dry weight of plants treated at flowering with 2.0 mM FA were reduced by 25% one week after treatment. Treatment with 2.0 mM FA at podfill caused the senescence and abscission of older leaves and reduced total plant leaf area, plant dry weight and mean pod dry weight by 54, 40, and 48%, respectively, one week after treatment. The plants treated at the seedling and flowering stages recovered by final harvest. In a subsequent experiment, FA (0, 0.50, 1.0, 1.5 mM) reduced total plant leaf area at the seedling and flowering stages but not at podfill. The youngest expanding leaves were most sensitive to FA at flowering. The leaf area of these leaves was reduced by 35 and 25%, one and two weeks after treatment, respectively. Their absolute growth rates were reduced from 31 to 56% one week after treatment at flowering. Their relative growth rates were reduced by 50% one week after treatment. Growth rates then recovered within two weeks after treatment. In the final experiment, biweekly exposures of FA (0.25, 0.50, 0.75, 1.0) reduced total plant leaf area but did not affect any other growth parameters. Weekly exposures of FA (0.25, 0.50, 0.75, 1.0) reduced total plant leaf area up to 34%, absolute growth rate up to 58%, leaf number up to 31% and pod number up to 58%. As the frequency of exposure to FA increased, the concentration necessary to affect bean plant growth and development decreased.     

The fission yeast mitotic activator cdc25 and sucrose induce early flowering synergistically in the day-neutral Nicotiana tabacum cv. Samsun

Here, the tobacco (Nicotiana tabacum) day-neutral (DN) cv. Samsun transformed with the Schizosaccharomyces pombe mitotic activator gene Spcdc25 was used to study the onset of flowering. Wild type (WT) and cdc25 plants were grown from seeds in vitro until they were 20 cm high. Apical and basal nodes were then subcultured repeatedly and the regenerated plants were used to document time to flowering and the number of leaves formed before flowering. Three sucrose treatments (3, 5 or 7% (weight/volume)) were used and measurements of leaf endogenous soluble carbohydrates were performed. In the 3% treatment, cdc25 plants flowered but WT plants did not. The higher sucrose treatments enabled WT flowering; two-thirds of the plants flowered at 5%, while all plants flowered at 7% sucrose. However, in all treatments, cdc25 plants exhibited significantly earlier flowering and fewer leaves compared with wild type. Remarkably, a typical acropetal flowering gradient in WT plants did not occur in cdc25 plants. In cdc25 leaves, there were significantly higher amounts of endogenous sugars with a higher proportion of sucrose compared with WT. Our data demonstrate that Spcdc25 expression and sucrose act synergistically to induce precocious flowering.     

Mutations in AP22.65 accelerate flowering in Arabidopsis thaliana

Identification of the gene(s) responsible for flowering time in Arabidopsis has significant implications. We used the T-DNA insertion library of Arabidopsis thaliana to screen an early-flowering mutant that exhibits accelerated flowering under short-day conditions. AP22.65, a novel flowering-time gene in that species, was isolated and identified via genome-walking and bioinformatics analysis. The flowering time of AP22.65-complementing plants was similar to that of the Col-0 wild type (WT). Conversely, its overexpression delayed flowering. Consistent with this phenotype, expression of AP22.65 was decreased in the ap22.65-1 mutant, recovered in AP22.65-complementing plants, and increased in AP22.65-overexpressing plants. Compared with the WT, expression levels of critical genes in different flowering pathways, i.e., SPY, FLC, GI, CO, FT, and LFY, were down-regulated in loss-of-function mutants. Expression of AP22.65 was distributed in flowers, siliques, rosette leaves, and whole seedlings. Therefore, this gene may be a negative regulator of Arabidopsis flowering.     

Overexpression of BpAP1 induces early flowering and produces dwarfism in Betula platyphylla × Betula pendula

The involvement of APETALA1 (AP1) in the flowering transition has been the focus of much research. Here, we produced Betula platyphylla × Betula pendula (birch) lines that overexpressed BpAP1 using Agrobacterium‐mediated transformation; we obtained five independent 35S::BpAP1 transgenic lines. Polymerase chain reaction (PCR), Southern, northern and western analyses were used to identify the transformants. As determined by quantitative real‐time PCR (qRT‐PCR), BpAP1 expression in roots, shoots, leaves and terminal buds of 35S::BpAP1 transgenic lines was significantly higher than that in the wild type (WT, P < 0.01). The average height of 2‐year‐old 35S::BpAP1 plants was significantly lower (41.17%) than that of non‐transgenic plants. In the 35S::BpAP1 lines, inflorescences emerged successively beginning 2 months after transplanting. In addition, the length–diameter ratio of fully developed male and female inflorescences were both significantly less than those of the WT (P < 0.05), i.e. the morphological characteristic was stubby. The male inflorescences emerged early, with empty, draped anthers, and pollen was rarely produced, whereas the female floret structure was not different from WT. The pistils developed normally and could accept pollen, leading to the production of hybrid progeny (F₁). F₁ plants completed flowering within only 1 year after sowing. We demonstrate that BpAP1 can be inherited through sexual reproduction. Overexpression of BpAP1 caused early flowering and dwarfism; these lines had an obviously shortened juvenile phase. These results greatly increase our understanding of the mechanisms underlying the flowering transition and enhance genetic studies of birch traits, and they open up new possibilities for the breeding of birch and other woody plants.     

Specification of Arabidopsis floral meristem identity by repression of flowering time genes

Flowering plants produce floral meristems in response to intrinsic and extrinsic flowering inductive signals. In Arabidopsis, the floral meristem identity genes LEAFY (LFY) and APETALA1 (AP1) are activated to play a pivotal role in specifying floral meristems during floral transition. We show here that the emerging floral meristems require AP1 to partly specify their floral identities by directly repressing a group of flowering time genes, including SHORT VEGETATIVE PHASE (SVP), AGAMOUS-LIKE 24 (AGL24) and SUPPRESSOR OF OVEREXPRESSION OF CO1 (SOC1). In wild-type plants, these flowering time genes are normally downregulated in emerging floral meristems. In the absence of AP1, these genes are ectopically expressed, transforming floral meristems into shoot meristems. By post-translational activation of an AP1-GR fusion protein and chromatin immunoprecipitation assays, we further demonstrate the repression of these flowering time genes by induced AP1 activity and in vivo AP1 binding to the cis-regulatory regions of these genes. These findings indicate that once AP1 is activated during the floral transition, it acts partly as a master repressor in floral meristems by directly suppressing the expression of flowering time genes, thus preventing the continuation of the shoot developmental program.     

The Suppression of WRKY44 by GIGANTEA-miR172 Pathway Is Involved in Drought Response of Arabidopsis thaliana

Water availability is an important environmental factor that controls flowering time. Many plants accelerate flowering under drought conditions, a phenomenon called drought escape. Four pathways are involved in controlling flowering time, but which ones participate in drought escape is not yet known. In this study, plants with loss-of-function mutations of GIGANTEA (GI) and CONSTANS (CO) exhibited abnormal drought-escape phenotypes. The peak mRNA levels of GI and FKF1 (Flavin-binding Kelch domain F box protein 1) and the mRNA levels of CO and FT (Flowering locus T) changed under drought stress. The microRNA factor miRNA172E was up-regulated by drought stress, and its up-regulation was dependent on GI, while other miRNA172s were not. Water-loss analyses indicated that gi mutants were more sensitive while miRNA172 over-expressing (miRNA172-OX) plants were less so to drought stress than wild-type plants. Digital gene expression and real-time PCR analyses showed that WRKY44 was down-regulated by GI and miRNA172. The WRKY44 protein could interact with TOE1 (a target of miRNA172) in a yeast two-hybrid system. We proposed that GI–miRNA172–WRKY44 may regulate drought escape and drought tolerance by affecting sugar signaling in Arabidopsis.     

Wheat SOC1 functions independently of WAP1/VRN1, an integrator of vernalization and photoperiod flowering promotion pathways

Heading time in bread wheat ( Triticum aestivum L.) is determined by three characters – vernalization requirement, photoperiodic sensitivity and narrow-sense earliness (earliness per se) – which are involved in the phase transition from vegetative to reproductive growth. The wheat APETALA1 ( AP1 )-like MADS-box gene, wheat AP1 ( WAP1 , identical with VRN1 ), has been identified as an integrator of vernalization and photoperiod flowering promotion pathways. A MADS-box gene, SUPPRESSOR OF OVEREXPRESSION OF CO 1 ( SOC1 ) is an integrator of flowering pathways in Arabidopsis . In this study, we isolated a wheat ortholog of SOC1 , wheat SOC1 ( WSOC1 ), and investigated its relationship to WAP1 in the flowering pathway. WSOC1 is expressed in young spikes but preferentially expressed in leaves. Expression starts before the phase transition and is maintained during the reproductive growth phase. Overexpression of WSOC1 in transgenic Arabidopsis plants caused early flowering under short-day conditions, suggesting that WSOC1 functions as a flowering activator in Arabidopsis . WSOC1 expression is affected neither by vernalization nor photoperiod, whereas it is induced by gibberellin at the seedling stage. Furthermore, WSOC1 is expressed in transgenic wheat plants in which WAP1 expression is cosuppressed. These findings indicate that WSOC1 acts in a pathway different from the WAP1 -related vernalization and photoperiod pathways.     

利用阵列分析水稻AP2/EREBP家族基因的表达特性

AP2／EREBP蛋白是广泛存在于高等植物中的且包含AP2／EREBP功能域的重要转录因子家族,通常可分为包含单功能域的EREBP类蛋白和包含两个功能域的AP2类蛋白,它们的功能涉及植物生长发育调控和对逆境应答等许多方面。据预测．水稻基因组编码150个左右的AP2／EREBP家族成员,但目前绝大多数蛋白的功能仍不清楚。为了解这些基因在水稻不同器官中的表达特性,我们以AP2／EREBP功能域的氨基酸序列为基础,从水稻基因组数据库中搜索到12个AP2类以及20个EREBP类预测基因,利用PCR扩增的编码区序列制备了这些预测基因的macro—array。以幼芽、幼根、幼叶、颖花和灌浆期成熟叶的cDNA为探针,杂交分析结果显示：不同AP2类预测基因之间的表达量差别较大,但同一个基因在不同器官中表达量基本一致：与此不同的是,大部分EREBP类预测基因在幼根和成熟叶片中表达量较高,而在幼芽和幼叶中表达量较低。这些预测基因的表达模式可能与它们的功能密切相关。