Transcriptome and morphological analyses of double flower formation in Dianthus chinensis

The double flower developmental process is regulated via a complex transcriptional regulatory network.To understand this highly dynamic and complex developmental process of Dianthus spp.,we performed a comparative analysis of floral morphology and transcriptome dynamics in simple flowers and double flowers.We found that the primordium of double flowers of‘X’was larger in size compared to that of simple flowers of‘L’in Dianthus chinensis.RNA-seq and Weighted Gene Co-expression Network Analysis(WGCNA)during flower development,identified stage-specific gene network modules.Expression analysis by RNA-seq indicated that a group of genes related to floral meristem identity,primordia position and polarity were highly expressed in double flowers genotypes compared to simple flowers genotypes,suggesting their roles in double-petal formation.A total of 21 DEGs related to petal number were identified between simple and double flowers.The experiments of in situ hybridization revealed that DcaAP2L,DcaLFY and DcaUFO genes were expressed in the intra-sepal boundary and petal boundary.We proposed a potential transcriptional regulatory network for simple and double flower development.This study provides novel insights into the molecular mechanism underlying double flower formation in Dianthus spp.

Reliable and Efficient Agrobacterium tumefaciens-Mediated Genetic Transformation of Dianthus spiculifolius

Dianthus spiculifolius is a perennial herbaceous flower with strong environmental adaptability and is an important ornamental ground cover plant.In this study,seeds of D.spiculifolius were used as explants for callus induction,adventitious bud differentiation,and rooting by adding different concentrations of 2,4-dichlorophenoxyacetic acid(2,4-D),6-benzyl aminopurine(6-BA),and naphthaleneacetic acid(NAA)to Murashige and Skoog medium.The calli generated were co-cultured with Agrobacterium tumefaciens EHA105 containing pBI121-GUS or pBI121-GFP plasmids for 30 min,and transgenic regenerated plants were obtained by kanamycin(30mg·L^−1)screening.RT-PCR confirmed the stable expression of the exogenous GUS and GFP genes in the D.spiculifolius.Theβ-glucuronidase(GUS)histochemical staining confirmed GUS gene expression in transgenic calli,adventitious buds,and regenerated plants of D.spiculifolius.The green fluorescent protein(GFP)visual analysis showed GFP gene expression in transgenic calli.Furthermore,subcellular localization analysis showed that the three organelle marker proteins were not only successfully expressed but also accurately localized to their corresponding organelles in D.spiculifolius callus cells.These results indicated a successful establishment of a reliable and efficient A.tumefaciens-mediated genetic transformation system,which will contribute to functional gene research and genetic improvement of D.spiculifolius.

A new cyclopeptide from Dianthus caryophyllus

A new cyclopeptide as well as a known cyclopeptide were isolated from the whole plants of Dianthus caryophyllus. Their structures were elucidated as cyclo （-Thr-Gly-Pro-Tyr-Phe-） （1） and cyclo （-Ala-Gly-Ser-IIe-Phe-Phe-） （2） by means of chemical and spectroscopic methods including MS, 1D and 2D NMR techniques.

Cytological Observation of Microsporogenesis in Male-Sterile Lines of Chinese Pink(Dianthus chinensis L.)

This study addressed the differences in microsporogenesis between male sterile and fertile lines of Chinese pink. The microsporogenesis processes of male sterile and fertile lines were histologically examined in squashed pollen grains and in paraffin embedded sections. A stable male-sterile line （H-37B） was obtained following six generations of inbreeding in a self-fertile line, followed by two generations of backcrossing. In the corresponding fertile line, development of the mature pollen grains was followed through the initiation of the sporogenous cell, microsporocyte formation, and the tetrad developmental period. In the male-sterile line, abortion of the developing pollen grains was observed to take place at various stages, namely, sporogenous cell growth, mother cell meiosis, and tetrad transformation to the uninuclear state. The pollen grains of the fertile line were spheroid, turgid, and viable. By contrast, the male-sterile line produced pollen that was irregular in shape, empty, and nonviable. The abortion of the microspore in the male-sterile line appeared to relate to abnormal growth of the tapetum layer.

High Frequency Plant Regeneration from Leaf Derived Callus of <i>Dianthus caryophyllus</i>L.

An efficient procedure was developed for in vitro callus induction, proliferation and regeneration of carnation cultivar (Dianthus caryophyllus L.) using leaf, nodal and inter-nodal explants on Murashige and Skoog’s medium (MS) supplemented with exogenous plant growth regulators. For morphogenic callus induction and proliferation from various explants, MS medium supplemented with 3.0 mg/l 2,4-D was highly efficient with 100% callus induction frequency from inter-nodal explants. Leaf explants showed quicker response than nodal and inter-modal explants, for callus initiation within 6 days of inoculation. Best grown callus was obtained from leaf explant. The leaf-derived callus was maintained up to several weeks, which indicated that 8-week incubation period was the most suitable for obtaining well proliferated, morphogenic callus. Temperature variation also affected the growth of in vitro induced morphogenic callus from various explants. Results have shown that 27°C proved to be the best temperature for morphogenic callus induction and proliferation from leaf and inter-nodal explants. Among the auxin-cytokinin combination, MS medium containing 1.0 mg/l N(6)-benzylaminopurin (BAP) and 2.0 mg/l NAA showed the highest efficiency of callus initiation and proliferation from leaf, nodal and inter-nodal explants. Light conditions proved better for callogenesis and proliferation from leaf, nodal and inter-nodal explants. Regeneration response from well grown morphogenic callus was prominent on MS medium supplemented with 3.0 mg/l BAP alone and 1.0 mg/l NAA with 3.0 mg/l BAP.

生长调节剂对香石竹(Dianthus caryophyllus)组培苗生根的影响

为了解不同生长调节剂及其浓度对香石竹组培苗生根培养的影响,通过在基本培养基内添加不同浓度的IAA和IBA进行组织培养试验并观察其生根和生长情况,来探索促进香石竹组培苗生根的最佳生长调节剂种类及其浓度。研究结果表明,10%次氯酸钠灭菌4 min时灭菌效果最佳;MS培养基中植株的根长与根数明显高于1/2MS和2MS培养基,而且生根较快,生根率高,植株长势好;1.0 mg/L的IAA和IBA都有利于根的伸长,并显著增加根数,其余浓度促进效果不显著,且浓度过高会抑制根的生长。各浓度IBA处理的植株长势均较好,其中1.0 mg/L的IBA处理的植株长势最好,最有利于香石竹组培苗生根培养。上述研究结果为香石竹切花离体快繁及工厂化生产提供进一步的理论依据。

瞿麦药学研究概况

在广泛文献检索基础上,对瞿麦的种属、鉴别、药理、临床应用及禁忌进行了综述,为进一步开发利用提供资料。

青霉素消除香石竹玻璃苗作用的研究

青霉素(Penicillin)是医学上广泛应用的低毒高效抗菌药物。近年来研究表明,它在高等植物代谢上具有多方面的作用。如能促进植物生长,利于插条产生不定根,增强核糖核酸的活性等。香石竹(Dianthus caryophyUus)试管苗在 MS+BA 2mg/L+NAA 0.2mg/L 培养基上增殖多代后,置于含青霉素10、20、40、80M·U/L(万单位)不同浓度的 MS 培养基上培养。30天后,在各浓度上均未观察到有玻璃苗发生。而在含 BA 2mg/L、MAA 0.2mg/L 的对照培养基上,

活性炭对香石竹试管苗继代培养的影响

[目的]研究活性炭对香石竹试管苗继代培养的影响，寻求最适香石竹试管苗增殖的活性炭用量，为组培苗培养基的改良提供一些有益的参考。[方法]将材料分别转接在活性炭浓度为0,2、4、6g／L的培养基中，观察不同浓度活性炭对香石竹试管苗芽增殖数量和速度、株高、节间距及玻化率的影响。[结果]当活性炭浓度在2～4g/L时，能显著提高试管苗的芽数、芽的增殖速率和株高，但对茎节段数的增加无明显作用。在培养基中加入活性炭可显著地降低试管苗的玻化率。[结论]活性炭对香石竹试管苗的增殖和降低玻璃苗有一定的作用。

海水胁迫对须苞石竹种子萌发及幼苗生长的影响

为比较须苞石竹(Dianthus barbatus L.)种子对盐碱的耐受性,利用1%、5%、10%、15%、20%、30%浓度的海水处理须苞石竹种子,观察其种子萌发和幼苗生长的情况。结果表明,须苞石竹种子的萌发参数各项指标以及幼苗根长、苗长均随着海水浓度的增加而有不同程度地降低。发芽率在海水浓度10%以内、发芽势与发芽指数在海水浓度5%以内、简化活力指数在海水浓度15%以内均与对照清水的差异不显著(P>0.05);根长在海水浓度20%以内、幼苗长在海水浓度5%以内与清水的差异不显著(P>0.05)。说明须苞石竹对低浓度海水具有一定的耐受性。

N-月桂酰乙醇胺对香石竹开放和衰老进程中花瓣微粒体膜组分和功能的调节

【目的】探讨N-月桂酰乙醇胺[N-lauroylethanolamine,NAE(12:0)]延缓香石竹切花衰老的作用机理。【方法】以5μmol·L^(-1)NAE(12﹕0)对香石竹切花(Dianthus caryophyllusL.)‘Red Barbara’进行瓶插处理,研究瓶插试验进程中NAE(12﹕0)对花瓣微粒体膜组分和功能的影响。【结果】香石竹切花开放和衰老进程伴随着花瓣微粒体膜磷脂含量、膜脂流动性、膜脂脂肪酸不饱和指数以及膜结合酶比活力的下降,外源NAE(12﹕0)处理能在切花盛开后期至初萎发生前明显滞缓香石竹花瓣微粒体膜上述指标的下降速率,同时降低作为膜衰老可靠指标的电导率的上升幅度。【结论】NAE(12:0)延缓香石竹切花衰老的作用机理与NAE(12﹕0)对花瓣微粒体膜磷脂降解以及膜脂脂肪酸组分的调节有关,由此延缓了花瓣微粒体膜脂流动性的下降速率,降低了膜渗漏的上升幅度,并维持了膜结合酶较高的比活力,从而推迟了切花初萎的发生。

香石竹茎尖分生组织培养的研究

对香石竹(Dianthus caryophyllus)进行了茎尖分生组织快速繁殖培养及其影响因素的研究。结果表明,大花香石竹比散枝香石竹接种成活率高,且以顶芽为外植体,在MS+6-BA0.5 mg/L+NAA0.4 mg/L上培养的丛芽分化率最高,达到87.5%。将顶芽、带腋芽的茎节、不带腋芽的茎节或茎段分别接种在MS+6-BA0.5 mg/L+NAA0.4 mg/L上培养,顶芽分化的芽数最多。光照12 h/d对香石竹丛芽生长有利。

磷酸钠对康乃馨切花保鲜效果的影响

以康乃馨(Dianthus caryophyllus L.)为材料,初步研究了不同磷酸钠浓度对康乃馨切花的观赏寿命、花径、鲜重等外观形态指标及水分平衡、花青素含量、过氧化氢酶(CAT)活性等生理指标的影响。结果表明,含有0.2 g/L的磷酸钠保鲜液可有效延长康乃馨切花寿命,促进切花鲜重的增加与花径的增大,有利于保持水分平衡,抑制花青素的降解,保持CAT酶活性。

不同保鲜剂对康乃馨切花保鲜效果的影响

以康乃馨(Dianthus cargophyllus)切花为试材,3%蔗糖、250 mg/L柠檬酸和50 mg/L 6-BA为基础保鲜液,分别添加150 mg/L 8-羟基喹啉(8-HQ)、15 g/L CaCl_2和100 mg/L AgNO_3,以蒸馏水作对照,研究不同试剂组合对康乃馨切花的保鲜效果。结果表明,AgNO_3处理能显著增加切花鲜重,维持切花水分平衡,并能有效维持切花鲜重,抑菌效果好,显著延长切花瓶插寿命,瓶插寿命可达19.7 d,是理想的保鲜液;其次为8-HQ处理,瓶插寿命为15.8 d,比对照处理增加81.6%;而CaCl_2处理不利于康乃馨切花保鲜。

环保型保鲜液对香石竹切花瓶插的保鲜效应

以蔗糖和8-羟基喹啉为保鲜液基本成分,研究不含银离子的瓶插保鲜液在分别添加6-苄氨基腺嘌呤(6-BA)、亚精胺(Spermidine,简称Spd)、维生素C和柠檬酸后对香石竹(Dianthus caryophyllus L.)切花的保鲜效果。结果表明,与去离子水对照相比,4种含有不同物质的保鲜液均能不同程度地增加香石竹切花的鲜重和花径,改善体内水分状况,延缓可溶性蛋白质的降解,降低丙二醛含量,延长瓶插寿命,其中以处理Ⅳ(30 g/L蔗糖+200 mg/L 8-羟基喹啉+30 mg/L 6-BA)效果最好,处理Ⅲ(30 g/L蔗糖+200mg/L 8-羟基喹啉+0.1 mmol/L Spd)次之,属于高效环保型保鲜剂。

香石竹栽培繁殖技术

随着我省人民生活消费水平的提高,对鲜花生产提出了更高的要求。各地相继出现了一些鲜花店。但货源多靠从上海、广州等地空运,使得鲜花的价格比外地高出许多,且不能做到周年供应。同时由于保鲜条件尚不具备,鲜花的质量也不能得到可靠的保证,从而限制了鲜花在我省的普及。因此,栽培和繁殖优良切花品种是在我省普及鲜花供应的一个重要措施。几年来,笔者对香石竹的生产、繁殖技术进行了大量的试验研究,结果表明:香石竹做为一种优良的切花品种,在我省有条件的地区可以大量生产,以填补我省鲜花生产的空白。

6-BA与NAA对黑玫瑰石竹离体培养的影响

以黑玫瑰石竹(Dianthus linn)茎段为外植体,MS培养基为基本培养基,研究了6-BA与NAA对黑玫瑰石竹试管苗不定芽分化的影响。结果表明,采用MS+1.0 mg/L 6-BA和0.2 mg/L NAA培养基对芽分化及生长效果较好。

切花香石竹标准化栽培技术

香石竹（Dianthus caryophyllus）即康乃馨,又名狮头石竹、麝香石竹、大花石竹、荷兰石竹。为石竹科石竹属类植物,现代切花香石竹品种是香石竹与其他种反复杂交并经由人工长期选择的结果,在温室里几乎可以连续不断开花。是传统四大切花之一,代表了健康和美好祝愿,也是母亲节的专门用花。扦插苗外观标准1.大小整齐一致。

Molecular Cloning and Characterization of Carnation EBF1 Gene During Flower Senescence and upon Ethylene Exposure and Sugar

A cDNA clone encoding a putative EBF-like protein （DCEBF1）was obtained from total RNA isolated from senescing carnation （Dianthus caryophyllus L.） petals using reverse transcription PCR and rapid-amplification of cDNA ends techniques. The cDNA contained an open reading frame of l 878 bp corresponding to 625 amino acids. Results of Northern blot indicated DCEBFI expression was enhanced by endogenous and exogenous ethylene, and was inhibited by STS in petals and ovaries. Upon wounding treatment, DCEBF1 showed a quick increase in mRNA accumulation which was positively correlated with the increase in ethylene production. The levels of DCEBF1 mRNA increased in both petals and ovaries by sucrose treatment compared with the control.

细胞分裂素处理对香石竹切花保鲜效果的影响

以香石竹切花为试材,通过测定其花径、瓶插寿命、鲜质量、水分平衡、可溶性蛋白质、相对电导率等指标,研究了6-苄氨基嘌呤(6-BA)、激动素(KT)、玉米素(ZT)3种细胞分裂素处理的瓶插液对香石竹切花保鲜效果的影响,以期筛选出理想的香石竹切花保鲜液。结果表明:在蒸馏水和可利鲜对照下,30 g·L-1 Suc(蔗糖)+250 mg·L-1 CA(柠檬酸)+200 mg·L-1 8-HQS(8-羟基喹啉硫酸盐)+10 mg·L-1 6-BA和30 g·L-1 Suc+250 mg·L-1 CA+200 mg·L-1 8-HQS+5 mg·L-1 ZT保鲜效果最佳。