Optimization of the uidA Gene Transfer of Rosa hybrida via Agrobacterium tumefaciens: an Assessment of Factors Influencing the Efficiency of Gene Transfer

To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing -glucuronidase gene. The results indicate that explant, light condition, salt concentration and acetosyringone (AS) concentration in co-culture medium are the most important factors, and factors like co-culture temperature, co-culture period and bacteria density have a strong effect on the growth of bacteria and then T-DNA transfer. Optimized co-cultivation was performed by inoculation of embryogenic callus with bacteria at a density of OD600= 0.50.8 for 20 min and co-culture in darkness under 23 C on medium with 1/2 MS salts and 300 mol稬1 AS for 3 d.

Agrobacterium tumefaciens-mediated transformation of modern rose(Rosa hybrida)using leaf-derived embryogenic callus

Rose(Rosa hybrida)is widely used for cut flowers and as garden plants.Stable and efficient transformation system is required for functional genomics of rose.Here,we established an efficient transformation method for rose using Agrobacterium tumefaciens-mediated transformation of embryogenic callus.Expanding rose leaves were used as explants to induce somatic embryos,which were subjected to transformation with A.tumefaciens strain GV3101 using Green Fluorescence Protein(GFP)as a marker gene.It took about 8 months to generate transgenic shoots from embryogenic callus.PCR,RT-PCR,Southern and Western blotting,as well as stereoscopic fluorescence microscopy analysis demonstrated that GFP transgenes integrated stably into the rose genome.According to our data,a transformation efficiency of up to 6%can be achieved by following this optimized protocol.

Direct Adventitious Bud Induction and Plant Regeneration of Rosa hybrida Samantha

Effect of explant, site of leaflet, induction period in the dark and combinations of plant growth regulators on direct adventitious bud induction and plant regeneration of Rosa hybrida Samantha was investigated. The results showed that after an induction period of 8 d on MS medium with 1.5 mg L-1 TDZ and 0.05 mg L-1 NAA in the dark and a subculture on MS medium with 0.5 mg L-1 BA and 0.01 mg L-1 NAA under light, the best plant regeneration was obtained and the regeneration frequencies of leaflets and petioles were 51.8 and 10% respectively. There was no significant difference in regeneration ability between leaflets at different sites of the compound leaves, longer time of induction in the dark or high concentration of auxin would cause callus formation, which was disadvantageous for shoot regeneration, and the regeneration frequency was significantly reduced. This regeneration system could be applied for genetic transformation of this cultivar in the future.

Exogenous ethylene influences flower opening of cut roses (Rosa hybrida) by regulating the genes encoding ethylene biosynthesis enzymes

The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, ‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process is inhibited by ethylene. Ethylene production and 1- aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment, ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage (stage 3) in ‘Samantha’, and they were much more dramatically enhanced and peaked at the later stage (stage 4) in ‘Kardinal’ than control during vasing. cDNA fragments of three Rh-ACSs and one Rh- ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-in- duced expression patterns of Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in ‘Kardinal’ than that of ‘Samantha’. As an ethylene action inhibitor, STS at concentration of 0.2 mmol/L generally inhib-ited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in ‘Kardinal’. Our results suggests that ‘Kardinal’ is more sensitive to ethylene than ‘Samantha’; and the changes of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in ‘Samantha’ and the inhibition in ‘Kardinal’. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding.

单瓣月季花Rosa chinensis var.spontanea的发现与历史

月季(Rosa hybrida L.)是世界最具经济价值的切花和园艺花卉,作为现代月季最重要的祖先之一——月季花(R.chinensis),一直以来都备受国内外育种家的高度关注。该文通过广泛查阅文献资料,对月季花及其野生型——单瓣月季花(R.chinensis var.spontanea Rehder & Wilson)的发现、发表和历史等多方面进行了阐述,拟为国内外月季育种家提供参考,为中国育种事业提供宝贵的理论依据和研究背景。

The MADS-box gene RhAGL6 plays a master role in regulating the receptacle malformation in rose at low temperature

Low temperature usually results in the developmental deformity of flower organs,immensely affecting the quality of rose flowers.However,it's largely unknown about the regulatory mechanisms activated by low temperature.Here,we used a low temperature-sensitive Rosa hybrida cv.‘Peach Avalanche’to screen a MADS-box gene RhAGL6 via conjoint analysis between RNA sequencing(RNA-seq)and whole-genome bisulfite sequencing(WGBS).Furthermore,we found that low temperature induced the hypermethylation and elevated histone 3 lys-27 trimethylation(H3K27me3)level on the RhAGL6 promoter,leading to decreased RhAGL6 expression.In addition,RhAGL6 silencing resulted in the formation of abnormal receptacles.We also found that the levels of gibberellins(GA3)and abscisic acid(ABA)in the receptacle under low temperature were lower and higher,respectively,than under normal temperature.Promoter activity analysis revealed that GA3 significantly activated RhAGL6 promoter activity,whereas ABA inhibited it.Thus,we propose that RhAGL6 regulates rose receptacle development by integrating epigenetic regulation and phytohormones signaling at low temperature.

Exogenous calcium enhances the physiological status and photosynthetic capacity of rose under drought stress

Drought(water shortage)can substantially limit the yield and economic value of rose plants(Rosa spp.).Here,we characterized the effect of exogenous calcium(Ca^(2+))on the antioxidant system and photosynthesis-related properties of rose under polyethylene glycol 6000(PEG6000)-induced drought stress.Chlorophyll levels,as well as leaf and root biomass,were significantly reduced by drought;drought also had a major effect on the enzymatic antioxidant system and increased concentrations of reactive oxygen species.Application of exogenous Ca^(2+)increased the net photosynthetic rate and stomatal conductance of leaves,enhanced water-use efficiency,and increased the length and width of stomata following exposure to drought.Organ-specific physiological responses were observed under different concentrations of Ca^(2+).Application of 5 mmol·L^(-1)Ca^(2+)promoted photosynthesis and antioxidant activity in the leaves,and application of 10 mmol·L^(-1)Ca^(2+)promoted antioxidant activity in the roots.Application of exogenous Ca^(2+)greatly enhanced the phenotype and photosynthetic capacity of potted rose plants following exposure to drought stress.Overall,our findings indicate that the application of exogenous Ca^(2+)enhances the drought resistance of roses by promoting physiological adaptation and that it could be used to aid the cultivation of rose plants.

非生物胁迫对月季生长发育影响的研究进展

从温度、光照、水分、盐及重金属5个非生物胁迫方面入手,对月季应答非生物胁迫的形态及生理生化变化、分子调控的研究现状进行综述,阐述非生物胁迫对月季生长发育影响,为深入阐释月季应答非生物胁迫的调控机制提供参考,也为高抗月季品种的培育和应用提供理论依据。提出相关研究应利用组学方法和先进的生物技术多维度解析月季应答非生物胁迫的分子调控网络,挖掘非生物胁迫应答关键基因,通过转基因技术培育抗性强的月季优势品种。

影响月季花瓣呈色的理化因子及花色苷组分分析

【目的】通过对影响月季花瓣呈色的各理化因子的定量评价及其相关性分析,探讨月季花色的形成机理,可为花色育种提供理论参考和受体品种,对于探究花色形成机理和种质创新具有重要意义。【方法】以8个不同花色的月季品种为试验材料,分别对其花瓣颜色参数、细胞液pH值、金属离子含量、总花色苷含量、总黄酮含量和总叶绿素含量等理化指标进行测定和比较,并对花色苷组分进行定量分析。【结果】(1)不同花色月季的理化因子间存在显著差异,其中细胞液pH值、Fe^(3+)、Ca^(2+)、Al^(3+)含量以及总花色苷、总黄酮含量等因子与花瓣颜色的形成密切相关,总花色苷含量和总黄酮含量的变化起直接作用,金属离子及细胞液pH值等因素通过改变花色素结构来影响花色。(2)不同花色月季花瓣中所含有的花色苷组分不同。其中,矢车菊素-3,5-葡萄糖苷在月季中占主体地位,主要调控紫红色花朵的形成;其次是天竺葵素-3,5-葡萄糖苷,其主要调控橙色、红色花朵的形成。黄色花朵中花色苷含量很少,主要受类胡萝卜素的调控;橙色花朵受花色苷和类胡萝卜素的双重调控;白色花朵中几乎不含有花色苷。芦丁在8个月季品种中均大量存在,但与花瓣形成无显著相关关系。(3)对8种月季花瓣中花色苷组分进行聚类分析时发现,粉紫色品种‘薰衣草花环’以极高的芦丁含量和少量的花色苷含量始终单独聚为一类。【结论】花色苷的种类和含量直接影响月季的花色,黄酮类物质则辅助花色苷的形成,而细胞液pH值、Fe^(3+)、Ca^(2+)、Al^(3+)含量等因子间接影响花色。在8个月季品种中,粉紫色品种‘薰衣草花环’以其优异的花色苷组分及较高的细胞液pH成为最适合进行花色改良的受体品种。

月季RhMAX2A基因的克隆与表达分析

【目的】克隆月季(Rosa hybrida L.)RhMAX2A基因c DNA序列,分析其序列特征及其在不同组织中和去顶后的表达情况,为探究该基因在月季中的生物学功能及调控侧枝发生的转导机制提供理论支持。【方法】以月季品种滇红(Rosa hybrida‘Dianhong’)为材料,通过RT-PCR技术克隆RhMAX2基因的cDNA序列,利用生物信息学方法对其序列和所编码的蛋白质进行分析,利用烟草(Nicotiana tabacum)瞬时转化技术分析蛋白的亚细胞定位,同时采用实时荧光定量PCR(qRT-PCR)检测其在不同组织中及去顶后的表达情况。【结果】RhMAX2A基因(GeneBank登录号为OP055810)cDNA序列长1030 bp,编码246个氨基酸,该蛋白分子式为C_(2910)H_(4793)N_(1029)O_(1244)S_(210),相对分子质量为27.35 kD,总原子量为3909;该蛋白不稳定系数为53.07,脂肪系数为106.30,GRAVY值为0.049,是一类不稳定亲水性蛋白;RhMAX2A蛋白的二级结构主要由α-螺旋和无规则卷曲构成,且RhMAX2A为推定的F-box结构域,属于α/β水解酶家族。同源序列比对和系统进化树关系分析结果表明,RhMAX2A氨基酸序列(OP055810)与同属的古老月季品种月月粉(Rosa chinensis‘Old Blush’)氨基酸序列(XP_024283944.1)相似性最高,其次是同亚科的草莓(Fragria vesca subsp.vesca)(XP_004287076.1),三者亲缘关系较近。亚细胞定位结果显示,RhMAX2A编码蛋白位于细胞核。qRT-PCR检测结果显示,RhMAX2A基因在根、腋芽和节中表达,根中表达量最高,去顶处理显著上调RhMAX2A基因在根和腋芽中的表达。【结论】成功克隆了滇红RhMAX2A基因,其编码蛋白在细胞核上发挥作用,主要在根和腋芽中表达,且受去顶诱导上调表达。

基于层次分析法的13个月季新品种性状综合评价

【目的】筛选出适宜在北京地区大量推广应用的优秀月季品种。【方法】对北京市园林绿化科学研究院自主培育的13个月季新品种,从生长性状、花部性状、抗病虫性三个方面,观测月季的株型、生长势、花量、花色、花径、四季开花性、抗白粉病、抗黑斑病等31个性状,采用层次分析法对性状进行综合评价。【结果】通过主成分分析筛选出17个重要性状,经综合打分并进行排序,13个月季新品种被分为两个等级,Ⅰ级有7个品种,分别为‘暗香’‘可爱冰淇淋’‘红五月’‘好风佳月’‘新时代’‘春花秋月’‘粉花毯’,Ⅱ级包括6个品种,分别为‘春潮’‘卢沟晓月’‘黄钻’‘闭月羞花’‘千纸鹤’‘东方之珠’。【结论】该评价体系为筛选适宜北京地区栽植的月季品种提供了依据,该研究筛选出的7个Ⅰ级月季品种适合在北京地区推广应用。

Study on the Introduction and Screening of Cut Roses in the Tropical Coastal Area of China

[Objective] This study aimed to select suitable cultivars of cut roses with prominent comprehensive performance which is suitable for the commercial promotion in the tropical region of South Hainan Province,including Sanya and the low-altitude area.[Method] Introduction experiment and production test were designed to observe the agronomic and economic traits of cultivars in a rose resources nursery established in Sanya.[Result] Through observation in the resources garden,18 varieties were chosen to conduct the introduction experiment,of which 8 varieties were selected to accomplish the production test.The results of the production test showed that ＇Vendela＇,＇Golden Emblem＇,＇Samantha＇ and ＇Tineke＇ showed the best adaptability;the cut flower quality of ＇Carola＇,＇Black Magic＇ and ＇Golden Emblem＇ belonged to the first grade;the yield of ＇Movie Star＇,＇Golden Emblem＇ and ＇Black Magic＇ was more than one million per hm2;except ＇Golden Emblem＇,the vase life of the other seven cultivars was more than 10 days;＇Carola＇,＇My Choice＇ and ＇Perfume White＇ had better disease-and insect-resistance;＇Carola＇,＇Vendela＇,＇Rouge Meilland＇,＇Tineke＇,＇Golden Emblem＇,＇Samantha＇,＇Perfume White＇,＇Saiun＇,＇Asagumo＇ and ＇Double Delight＇ were more resistant to blackspot,while ＇Black Magic＇,＇Versilla＇,＇Alec＇s Red＇ and ＇Movie Star＇ were less resistant to blackspot.[Conclusion] Cultivars of cut roses were able to grow and develop normally in the tropical coastal area of Hainan Province,including South Hainan and Sanya,where it was appropriate for the production of cut roses.However,it is necessary to pay attention to the prevention and control of thrips and blackspot.＇Black Magic＇,＇Carola＇,＇Vendela＇,＇Rouge Meilland＇,＇Golden Emblem＇,＇Tineke＇,＇Movie Star＇ and ＇Samantha＇ showed best comprehensive performance,which were thus worthy of large-scale popularization in Hainan Province.

The Waterlogging Tolerance of Cut Rose by Field Observation and Variety Selection in Sanya

[Objective] The aim was to master waterlogging tolerance of cut rose vari-eties, and provided technical reference for promoting cut rose largely. [Method] Based on natural rainfal information from July to October in Sanya, comprehensive performance of 22 varieties were observed in two consecutive years to analyze wa-terlogging tolerance ability of cut rose at waterlogging disaster. [Result] The mortality rates of Diana, Black Magic, Eric Red and Vendela were over 15%, and Diana ’s was 25.7%. Carola, Rouge Meil and, Perfume white, Lovers ’ Meeting, Choice, Tineke, Vendela, Marina, Samantha, Golden Emblem, Asagumo, Pink Fan and Dou-ble Delight grew better and recovered quickly after the disaster, with waterlogging tolerance. [Conclusion] The waterlogging tolerance of Carola, Rouge Meil and, My Choice, Tineke, Vendela, Marina, Samantha, Golden Emblem, Asagumo, Pink Fan and Double Delight was the best, which is also true for their comprehensive perfor-mances. But the waterlogging tolerance of other major cultivars such as Black Magic, Movie Star, Tineke and Vendela was poorer.

The Expression of RcLEA Gene Improves Tolerance of E. coli Cells to Abiotic Stress

[Objective] This study was to reveal the heat induced expression model of RcLEA gene and its tolerance to various abiotic stresses.[Method] Heat resistant and heat sensitive varieties of Rosa hybrida L.were subjected to heat shock treatment at 38 ℃ for 3 h;then RcLEA gene from both varieties treated was cloned and transformed into Escherichia coli strain BL21;finally recombinant colonies were separately cultured at 4 ℃ and 50 ℃ under the stresses of LiCl,NaCl,Na2CO3,CdCl2 and H2O2 to study the responses of recombinant E.coli strains to high temperature,low temperature and some other abiotic stresses.[Result] After heat shock treatment at 38 ℃ for 3 h,RcLEA gene expressed highly in ＇Schloss mannieim＇（SM）and ＇Las vegas＇（LV）variety,but weakly or even not expressed in ＇Kordes＇ Perfecta＇（KP）,indicating that this gene is closely related with heat resistance of R.hybrida.Compared with WT strains,recombinant clones showed higher tolerance to abiotic stresses including high temperature,low temperature,heavy metal,high salt,high pH value and oxidation,suggesting that RcLEA is concerned with the response of R.hybrida to abiotic stresses mentioned above.[Conclusion] These results provide thoughts for increasing heat resistance by introducing RcLEA into heat sensitive R.hybrida varieties and studying the heat-resistant mechanism of R.hybrida,and also provide theoretical support for selecting heat resistant variety of landscape and ornamental plants like R.hybrida.

月季高温胁迫研究进展

为推动耐热月季种质资源筛选和育种进程,总结了高温胁迫对月季形态、产量、品质、生理生化(细胞膜透性、渗透调节物质含量、抗氧化酶活性)的影响,简述了月季对高温胁迫响应机理的研究现状,阐述了月季耐热性评价指标筛选、种质资源耐热性评价以及耐热月季选育工作进展。结果表明:相较于其它花卉,月季研究目前集中于种质资源耐热性评价,而高温响应机理和耐热品种选育则相对滞后。随着全球气候变暖,高温胁迫已是现今月季面临的重要逆境胁迫。目前,亟需结合多组学深入研究其相应机理,以加快耐热月季种质筛选和育种进程,以满足华东地区对耐热月季品种的需求。

月季花青素苷相关R_2R_3-MYB蛋白基因的克隆和表达分析

【目的】花青素苷是月季花瓣呈红色或粉色的重要因素。R2R3-MYB蛋白是调控植物花青素苷合成的关键转录因子。从月季花瓣中克隆花青素苷调控相关的R2R3-MYB蛋白同源基因,并分析这些基因与月季花瓣颜色和花青素苷合成的关系,为花色基因工程改良奠定基础。【方法】根据植物花青素苷调控相关R2R3-MYB蛋白的保守序列设计简并引物,结合3'-RACE和Y-RACE方法从月季花瓣中扩增同源基因的全长编码序列。用植物第四(Sg4)和第六(Sg6)亚家族的R2R3-MYB蛋白、拟南芥次生代谢调控相关的R2R3-MYB序列和克隆基因的编码蛋白进行多序列比较和进化分析。通过比较不同颜色的月季花瓣中花青素苷含量和R2R3-MYB蛋白基因的表达水平,分析月季花瓣中R2R3-MYB蛋白基因与花青素苷调控的关系。【结果】从月季‘红胜利’的红色花瓣中克隆了2个R2R3-MYB蛋白基因(Rh MYBs4-1和Rh MYBs6-1,Gen Bank登录号分别为KJ664810和KJ664811)。序列分析表明,Rh MYBs4-1和Rh MYBs6-1蛋白均含有保守的R2R3-MYB结构域,分别与植物Sg4和Sg6 R2R3-MYB蛋白同源。Rh MYBs4-1具有Sg4 MYB蛋白典型的C1、C2抑制子和锌指结构。Rh MYBs6-1具有Sg6 MYB蛋白特有的(A/S/G)NDV和KPRPR(T/S)基序。表达分析显示Rh MYBs4-1和Rh MYBs6-1均在月季‘红胜利’的花瓣中高水平表达,在叶片和花药中表达水平很低。比较7种不同颜色月季花瓣中的花青素苷含量和Rh MYBs4-1和Rh MYBs6-1的表达水平,发现红色花瓣中花青素苷含量远高于其他材料,相应地,Rh MYBs4-1和Rh MYBs6-1均在红色花瓣中具有最高的表达水平。在粉红色月季花瓣中,花青素苷含量不到红色花瓣的10%,Rh MYBs4-1的表达水平极低,而Rh MYBs6-1的表达水平与红色花瓣相当。【结论】月季Rh MYBs4-1和Rh MYBs6-1分别编码Sg4和Sg6亚家族的R2R3-MYB蛋白,均在红色月季花瓣中高水平表达,可能是月季花青素苷合成和花瓣颜色的重要调控基因。

不同配比保鲜剂对月季切花保鲜效果研究初报

以GA3为主剂,研究了不同配方保鲜剂对月季切花瓶插寿命及其水分平衡值等相关指标的影响。结果表明,与对照相比,处理H(10 g／L蔗糖+200 mg／L 8-HQ+100 mg／L GA3)极显著延长了月季切花瓶插寿命 (延长2．01 d)和盛开时间(延长2．80 d),花径增大1．95 cm(P<0．01),明显延缓了水分平衡值的下降及花枝鲜重变化率零值的到来,其各项指标均优于其他配方,为最优保鲜剂。

切花月季Samantha失水胁迫耐性与其超氧化物歧化酶之间的关联

以探讨月季切花失水胁迫耐性与其超氧化物歧化酶(SOD)之间的关联为目的,选用耐失水胁迫品种Samantha,用DDTC(SOD专一性抑制剂)进行12h预处理,之后在20℃、RH35%～40%、80μEm-2·s-1光照(12h/d)条件下进行12～48h的失水胁迫处理,然后进行瓶插观察。结果表明,20mmol·L-1DDTC预处理在显著抑制花瓣SOD活性的同时,明显减弱切花失水胁迫耐性,表现在切花弯颈率明显提高,复水率和水势值明显降低,瓶插寿命显著缩短。在花朵自然开放初期,花瓣MnSOD、FeSOD和Cu/ZnSOD这3种类型都存在。其中,MnSOD有1种,FeSOD有3种,Cu/ZnSOD有2种;迁移率大小依次为Cu/ZnSOD>FeSOD>MnSOD;谱带强弱依次为FeSOD>Cu/ZnSOD>MnSOD。经失水胁迫处理花瓣中诱导产生了3条新的FeSOD(FeSOD4、FeSOD5和FeSOD6)条带,与此同时MnSOD条带消失。DDTC预处理抑制了任何与上述失水胁迫诱导有关的新的条带产生。由此推测,切花月季Samantha失水胁迫耐性与其SOD活性相关,与FeSOD4～6的诱导产生相关联。

月季花发育过程中花色变化的生理生化研究

以月季‘仙境’为材料,观测其正面和反面花瓣在开花过程中6个阶段的颜色和其花色的L*、a*、b*、C*、h*值变化,花瓣组织结构,以及pH值、花色素含量、类黄酮含量、可溶性糖和可溶性蛋白含量,探讨月季花色的呈色机理。结果显示:(1)半开期时,花瓣正面红色最深,花色苷、类黄酮、可溶性糖含量最高,而可溶性蛋白含量则在盛开期时达到最高。(2)pH越小时,花瓣红色越深,而且pH值小范围内的变动,就能导致花色的改变。(3)花色素大部分集中分布在上表皮,并且上表皮呈圆锥形小突起、下表皮为扁平状,故上表皮花色深于下表皮,亮度小于下表皮。(4)a*正面与pH值、花色苷和可溶性糖含量呈正相关关系;a*反面与L*正面、L*反面、b*反面呈正相关关系;L*正面与b*正面、L*反面与b*反面呈极显著负相关关系。(5)众多影响花色呈现的因素中,正面(反面)花色主要是受到花色素苷(L*反面)的直接作用,其他因素则通过影响花色素苷(L*反面)间接影响花色的呈现。

微型月季品种分类的花粉形态学

【目的】明晰微型月季花粉形态特征与其品种分类关系。【方法】利用光学显微镜和扫描电子显微镜观察10个微型月季品种花粉形态特征,并根据花粉形状、瘤状纹饰密度、网眼密度和条纹密度等性状对不同微型月季品种进行聚类分析。【结果】(1)微型月季花粉具有3个萌发孔,极面观为三裂圆形;花粉外壁纹饰为瘤状雕纹、网眼、条纹的不同组合,不同品种外壁纹饰差异较大;根据极轴/赤道轴值(P/E值),微型月季花粉可分为长球形和超长球形。(2)聚类结果表明,在相似系数为0.37时,可分为3组:第1组包括‘白柯斯特’、‘粉柯斯特’、‘紫微星’和‘罗曼蒂克’,特点是花粉外壁纹饰的条纹、网眼均密集;第2组包括‘金太阳’、‘小女孩’、‘矮仙女’、‘草裙舞女’和‘冬梅’,特点是花粉外壁纹饰的条纹密集,具瘤状雕纹,无网眼或较少;第3组为‘红云拉’,特点是花粉外壁纹饰的条纹和网眼较少。(3)结合不同微型月季品种外部形态指标的比较分析表明,孢粉学分类结果与形态学分类结果相似,但并不完全一致。【结论】不同微型月季品种花粉形状、萌发沟等特征近似,外壁纹饰差异较大,孢粉学分析结果可作为其品种分类依据,且分类结果总体支持形态学分类结果。