ISSR Analysis of Genetic Relationship of Germplasm Resource in Prunus mume Sibe. et Zucc

Using ISSR technique to analyze the diversity and genetic relationship of germplasm resources in 39 Prunus mume Sibe. et Zucc., the result showed that 10 primers were screened with high resolution from 51 primers, 120 fragments were amplified, of which 98 were polymorphic loci, accounting for 81.67% of total. Tested materials were divided into 3 classes, as was fundamentally accorded with the traditional classification base on horticulture. There was no obvious difference in geographic relationship among the clustering results.

梅(Prunus mume)IRAP分子标记技术体系的建立

基于梅Ty1-copia类逆转座子的长末端重复序列信息设计IRAP引物,通过五因素四水平L16(45)的完全随机正交试验、不同浓度PAGE胶等的研究,建立了适合梅IRAP标记的技术体系。结果表明,体系总体积25μL,其中基因组DNA 30 ng、25 mmol·L-1 MgCl2 2.0μL、2.5 mmol·L-1 dNTPs 2.5μL、10×PCR Buffer 2.5μL、Taq DNA聚合酶1.0 U、10μmol·L-1 LTR引物1.0μL,加ddH2O至25μL为最优IRAP的PCR反应体系,以及6.0%和8.0%浓度的非变性PAGE胶均适于IRAP多态性检测。这为进一步利用IRAP分子标记研究梅的遗传多样性和亲缘关系等奠定坚实的基础。

关于梅花Prunus mume的品种分类体系

探讨了在编著《中国梅花品种图志.中英双语增订版》时,为解决与国际接轨而遇到的先决问题。经钻研《国际栽培植物命名法规》(第7版)后,在梅种之下将品种分为3个品种群,即:①真梅品种群True Mume Group,②杏梅品种群Apricot Mei Group,③樱李梅品种群Blireiana Group。品种群以下,则不再分类、型等级别。而将二元分类法的内涵,融入品种名称之中。不同意樱李梅用独立的杂种学名Prunus×blireiana,也不同意把杏梅提升为独立的种名Prunus×bungo。樱李梅和杏梅还都应列在梅种之下,因三者之内果皮表面均具蜂窝状点穴——此乃梅、杏、李鉴别分类中的最关键性状,必须首先考虑,不能与其他性状等量齐观。

Advanced hepatocellular carcinoma responds to MK615, a compound extract from the Japanese apricot “Prunus mume ”

MK615, a compound extracted from the Japanese apricot "Prunus mume " has been reported to have in vitro anti-tumor activities against several cancer cell lines,including hepatocellular carcinoma(HCC). However, the clinical effects and feasibility of administering MK615for patients with HCC were unknown. We experienced a case with advanced HCC for which MK615 was effective against both lymph node and pulmonary metastases. A 60-year-old female underwent surgical resection of a 9 cm HCC in the right lobe. The pathological diagnosis was moderately differentiated HCC with vascular invasion. The HCC recurred in the liver 8 mo after the surgery. Radiofrequency ablation and transarterial infusion chemotherapy were performed, but the recurrence was not controlled. One year after the intrahepatic recurrence, pulmonary and lymph metastasis appeared.Sorafenib was administered, but was not effective.Then, MK615 was administered as a final alternative therapy after informed consent was obtained from the patient. Three months later, her alpha-fetoprotein level decrease and both the lymph node and pulmonary metastases decreased in size. The patient has survived for more than 17 mo after the MK615 administration, and was in good condition. Although further investigations are necessary to clarify its safety and efficacy in humans, MK615 may be useful for the treatment of HCC,without serious adverse effects.

New anti-proliferative agent,MK615,from Japanese apricot “Prunus mume” induces striking autophagy in colon cancer cells in vitro

AIM: To investigate the anti-neoplastic effects of MK615, an extract from the Japanese apricot (Prunus mume), against colon cancer cells. METHODS: Three colon cancer cell lines, SW480, COLO, and WiDr, were cultured with MK615. Growth inhibition was evaluated by cell proliferation assay and killing activity was determined by lactate dehydrogenase assay. Induction of apoptosis was evaluated by annexin Ⅴ flow cytometry. Morphological changes were studied by light and electron microscopy, and immunofluorescence staining with Atg8. RESULTS: MK615 inhibited growth and lysed SW480, COLO and WiDr cells in a dose-dependent manner. Annexin Ⅴ flow cytometry showed that MK615 induced apoptosis after 6 h incubation, at which point the occurrence of apoptotic cells was 68.0%, 65.7% and 64.7% for SW480, COLO, and WiDr cells, respectively. Light and electron microscopy, and immunofluorescence staining with Atg8 revealed that MK615 induced massive cytoplasmic vacuoles (autophagosomes) in all three cell lines. CONCLUSION: MK615 has an anti-neoplastic effect against colon cancer cells. The effect may be exerted by induction of apoptosis and autophagy.

创建“梅(Prunus mume)品种国际登录精品园”有感——陈俊愉教授与梅花

著名梅专家陈俊愉教授是梅品种国际登录权威。介绍了陈教授对梅花研究和梅事业的杰出贡献以及创建北京“梅品种国际登录精品园”的重大意义。

Photoperiod-and temperature-mediated control of the ethylene response and winter dormancy induction in Prunus mume

Plant dormancy is essential for perennial plant survival.Different genotypes of Prunus mume,including Eumume group and Apricot Mei group,undergo leaf senescence and dormancy at different times.In order to verify the cold resistance of P.mume,freeze resistance evaluation was carried out.Our results showed that Apricot Mei group had a stronger freezing tolerance than Eumume group and that leaf senescence and dormancy of Apricot Mei group occurred at an earlier period before winter.Based on phenotypic data in response to seasonal climate change,the significant candidate regions were selected using GWAS.Furthermore,through KEGG pathway and qRT-PCR analyses,we found that the ethylene-related genes,including PmEIL(Pm002057)and PmERF(Pm004265),were significantly upregulated in‘Songchun’Mei(Apricot Mei group)and downregulated in‘Zaohua Lve’Mei(Eumume group).Ethylene-related genes expression models showed that ethylene may be indirectly involved in the induction of dormancy.The PmEIL and PmERF genes were the core genes of the ethylene signal transduction pathway and were regulated by the exogenous ACC or PZA compounds.For non-dormant or weekly dormant perennial plants,application of ACC was able to induce plant dormancy and thus enhance cold/freeze tolerance.Overall,the expression of the major ethylene genes played a significant role in dormancy induction and freezing tolerance in P.mume;accordingly,application of ACC and PZA compounds were an effective approach for enhancing cold/freeze of tolerance of woody plant.

Genetic Relationships among Prunus mume var. pendula Using AFLP Markers

Genetic relationships among Prunus mume var. pendula were studied by using AFLP markers. 18 accessions representing 14 cultivars of Prunus mume var. pendula were selected from the germplasm collection at the Research Center of China Mei Flower. Seven Mse I-EcoR I AFLP primer combinations revealed 450 legible bands, and 269 of which were polymorphic markers. A similarity matrix was prepared using the simple matching coefficient of similarity and Neis (72) distance coefficient. A UPGMA dendrogram demonstrated the genetic relationships of the cultivars. The information given by AFLP markers was basically consistent with the morphological classification and the evolutionary history of the morphotypes, and roughly supported the new revised classification system for Chinese Mei Cultivars. But there were still several exceptions: 1) the Guhong Chuizhi inserted between the Tiaoxue Chuizhi and the Danfen Chuizhi; 2) the Wufu Chuizhi kept off the Pink Pendant Form, and the Moshan Chuizhi was removed from Viridiflora Pendant Form; 3) the Danbi Chuizhi and the Shuangbi Chuizhi of Viridiflora Pendant Form got together well but fell within the Pink Pendant Form.

Genome-wide identification and characterization of the bHLH gene family in an ornamental woody plant Prunus mume

The basic helix-loop-helix(bHLH)transcription factor family is the second-largest family in plants,where it plays essential roles in development,and the responses to multiple abiotic and biotic stressors.However,little information is available about this gene family in Prunus mume,which is widely cultivated in East Asia as an ornamental fruit tree.Here,100 PmbHLH genes were identified,and their evolution and functions were explored in P.mume for the first time.The PmbHLH genes were classified into 21 subfamilies.The chromosomal distribution,physicochemical properties,bHLH domain,conserved motif,and intron/exon compositions were also analyzed.Furthermore,the evolutionary pattern,divergence time of the PmbHLH family,and genetic relationships among P.mume,Arabidopsis thaliana,and Prunus persica and Fragaria vesca of Rosaceae were explored.The functional prediction analysis of these PmbHLHs indicated that their functions varied,and included participating in the formation of organs and tissues,responding to stress,and the biosynthesis and metabolism of hormones and other secondary metabolites.Interestingly,expression analyses of PmbHLHs also revealed diverse expression patterns.Most of the PmbHLH genes were highly expressed in roots and stems,and a few were highly expressed in leaves,buds,and fruits,indicating tissue expression specificity.Eight PmbHLH genes,which were upregulated during low-temperature stress,may have critical roles in the response to cold stress.Ten PmbHLHs were differentially expressed between weeping and upright branches in a P.mume F_(1) population.These results shed light on the structure and evolution of the PmbHLH gene family,and lay a foundation for further functional studies of the bHLH genes.

Inhibitory Activity of Prunus mume,Copitis chinensis and Crataegus pinnatifida on Vibrio harveyi and Its Biofilm

[Objective] This study was conducted to determine the in-vitro inhibitory effects of Prunus mume,Coptis chinensis and Crataegus pinnatifida on Vibrio harveyi and its biofilm. [Method]The inhibitory zone diameters of the three Chinese herbal medicines against V. harveyi were determined by agar diffusion method; the minimal inhibitory concentration( MIC) and minimal bactericidal concentration( MBC) values of the three Chinese herbal medicines against V. harveyi were determined by doubling dilution method; and the effects of the three Chinese herbal medicines on the formation of V. harveyi biofilm were determined by methyl thiazolyl tetrazolium( MTT) method. [Result]The three Chinese herbal medicines all inhibited V. harveyi to different degrees. C. chinensis and C. pinnatifida and P. mume exhibited the inhibitory zone diameters of( 17. 62 ± 0. 04),( 20. 16 ± 0. 08) and( 30. 76 ± 0. 26) mm against V. harveyi,respectively. P. mume and C. pinnatifida had strong inhibitory effects on V. harveyi. The MIC and MBC values of P. mume against V. harveyi were 7. 812 5 mg/ml; the MIC and MBC values of C. pinnatifida against V. harveyi were 31. 25 mg/ml; and the MIC and MBC values of C. chinensis against V. harveyi were 62. 5 mg/ml. P. mume had the strongest antibacterial and bactericidal ability. The MIC values of C. pinnatifida,C. chinensis and P. mume against V. harveyi were 7. 81,7. 81 and 1. 96 mg/ml,respectively,i. e.,P. mume exhibited the lowest MIC. [Conclusion] P. mume,C. pinnatifida and C. chinensis all have inhibitory effects on V. harveyi and its biofilm,and P. mume has the strongest bactericidal ability.

The Use of Prunus mume in the Development of Low Chill Apricot Varieties:the Status of Work in the United States

Several groups in the USA have been working to de-velop a low chill apricot that would ripen its fruits veryearly in the season. Unfortunately, the cultivatcd apricothas been developed mainly from the high chilling Euro-pean group within the species Prunus armeniaca. Amongthe five groups within the apricot species, this is the mostrecently evolved and least variable (Bailey and

Orthogonal Test Optimization of Extraction Technology of Total Fla-vonoids from Fruits and Leaves of Prunus mume Sieb. et Zucc. and Its in Vitro Anti-oxidation Activity

[Objectives]To optimize the extraction technology of total flavonoids component,and investigate its in vitro anti-oxidation activity.[Methods]The single factor was inspected firstly. By orthogonal experiment,the best extraction conditions of total flavonoids from fruits and leaves of P. mume Sieb. et Zucc. were determined,and reducing ability of the extracted total flavonoids and its DPPH and ABTS scavenging abilities were explored. [Results] The best extraction technology conditions: solid-liquid ratio of 1∶ 50,ethanol concentration of 50%,extraction time of 2. 5 h,extraction temperature of 85 ℃,two-time extraction. By detecting DPPH and ABTS scavenging abilities of total flavonoids,the anti-oxidation activity of the total flavonoids from fruits and leaves of P. mume Sieb. et Zucc. was analyzed and evaluated. [Conclusions]Fruits and leaves of P. mume Sieb. et Zucc. had a certain in vitro anti-oxidation activity,and heat reflux extraction method of its total flavonoids had high extraction rate and simple and convenient operation,which had some practical value.

Comparative Analysis of Fingerprint Spectra of Fruits and Leaves of Prunus mume Collected at Various Altitudes in Different Months

[Objectives] To study characteristic fingerprint spectra of fruits( its stones were removed) and leaves of Prunus mume collected at various altitudes in different regions of Jinchuan County,the Aba Tibetan and Qiang Autonomous Prefecture,Sichuan Province,China in different months by using Ultra Performance Liquid Chromatography( UPLC).[Methods]The evaluation system of similarity of chromatographic fingerprint spectra of traditional Chinese medicine and SPSS19. 0 were used to assess their similarity and to establish the reasonable reliable common model of chromatographic fingerprints,and the quality of the medicinal materials was evaluated.[Results] Seen from the fingerprint spectra of fruits( its stones were removed) and leaves of P. mume collected at different altitudes in various months,there were 15 common peaks in the fingerprint spectra of the fruits( its stones were removed),and the similarity of the 28 samples was higher than 0. 900,showing that there were small differences between the samples of the fruits( its stones were removed) in chemical components; there were 10 common peaks in the fingerprint spectra of the leaves,and the similarity of the 28 samples was also higher than 0. 900,indicating that there were also small differences between the samples of the leaves in chemical components.[Conclusions] The method of using fingerprint spectra to control the quality of fruits and leaves of P. mume was simple and stable and had strong specificity and good repeatability.

Survey of Annual and Seasonal Fungal Communities in Japanese <i>Prunus mume</i>Orchard Soil by Next-Generation Sequencing

Fungi play a vital role in the management of soil environment. Although various fungal communities are found in soil, it is difficult to determine the fungal community structure in soil. In this study, we conducted a comprehensive survey of fungal communities in Japanese Prunus mume orchard soil from 2010 to 2012 growing seasons using next-generation sequencing technology. Fungal DNA was directly extracted from the soil samples and the internal transcribed spacer 1 region was amplified by PCR and sequenced. We identified 34,826 fungal clone sequences from the soil samples. The fungal clones were sorted into 2132 operational taxonomic units and a majority of the discriminated clone sequences were classified as Ascomycota and Basidiomycota. The number of fungal species belonging to Ascomycota showed increases in June in the three growing seasons. That belonging to Glomeromycota showed increases in August in the three growing seasons. As Ascomycota fungi are wood decomposers and saprotrophs, the results suggested that the number of plant pathogenic fungi increased in Japanese P. mume orchard soil in June. These findings show for the first time the annual and seasonal fungal community structures in Japanese P. mume orchard soil, and are expected to provide valuable clues for improvement when planting new P. mume trees in Japanese orchards.

Inhibitory Effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio parahaemolyticus and Its Biofilm in Vitro

In order to explore the inhibitory effects of Prunus mume,Crataegus pinnatifida and Coptis chinensis on Vibrio parahaemolyticus and its biofilm in vitro,the agar diffusion method was applied. These three Chinese herbal medicines had different inhibitory effects on V. parahaemolyticus. The inhibition zone of C. pinnatifida to V. parahaemolyticus was( 15. 25 ± 0. 53) mm,and the minimum inhibitory concentration( MIC) and minimum bactericidal concentration( MBC) of C. pinnatifida on V. parahaemolyticus were both 31. 25 mg/m L; the inhibition zone of C. chinensis to V. parahaemolyticus was( 18. 08 ± 0. 10) mm,and the MIC and MBC of C. chinensis on V. parahaemolyticus were both15. 63 mg/m L; the inhibition zone of P. mume to V. parahaemolyticus was( 28. 99 ± 0. 47) mm,and the MIC and MBC of P. mume on V. parahaemolyticus were both 7. 81 mg/m L. The effects of three traditional Chinese medicines on the biofilm formation of V. parahaemolyticus were tested by MTT colorimetric method using methylthiazolyl tetrazolium( MTT). P. mume,C. pinnatifida and C. chinensis have significant inhibitory effects on V. parahaemolyticus biofilm and their MIC are 7. 81 mg/m L,3. 125 mg/m L,and 62. 5 mg/m L,respectively( P < 0. 01).The experimental results are expected to provide certain references for the development of new fishery drugs.

Inhibitory Effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio alginolyticus and Its Biofilm In Vitro

[Objectives]To explore the inhibitory effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio alginolyticus and its biofilm in vitro. [Methods] The agar diffusion method was applied to determine the inhibition zone diameter of three Chinese herbal medicines on V. alginolyticus; double dilution method was used to determine the minimum inhibitory concentration( MIC) and minimum bactericidal concentration( MBC) of three Chinese herbal medicines on V. alginolyticus; methylthiazolyl tetrazolium( MTT) method was applied to evaluate the effects of three Chinese herbal medicines on the biofilm formation of V. alginolyticus. [Results]The three Chinese herbal medicines had different inhibitory effects on V. alginolyticus. The inhibition zones of P. mume,C. chinensis,and C. pinnatifida on V. alginolyticus were( 18. 31 ± 0. 29) mm,( 20. 31 ± 0. 30) mm,and( 30. 24 ± 0. 14) mm respectively,and P. mume had strong bacteriostatic effects on V. alginolyticus. The double dilution method was used to determine the bacteriostatic effects of three Chinese herbal medicines on V. alginolyticus. Results showed that P. mume has the highest antibacterial and bactericidal ability. The MIC and MBC of P. mume,C. pinnatifida,and C. chinensis on V. alginolyticus was 3. 91,31. 25,and 125 mg/m L,respectively. Finally,MTT method was applied to evaluate the effects of C. pinnatifida,C. chinensis,and P. mume on the biofilm formation of V. alginolyticus,and results showed that the MIC on the biofilm formation of V. alginolyticus was 15. 63,7. 81,and 3. 91 mg/m L,respectively; the MIC of P. mume was the lowest. [Conclusions]This experiment proves that P. mume,C. chinensis,and C. pinnatifida have inhibitory effects on V. alginolyticus and its biofilm formation. It is expected to provide references for prevention and control of pathogenic vibrio induced diseases of aquatic animals.

梅子中感官活性成分的研究

应用HP-20大孔树脂柱层析、半制备高效液相色谱等多种分离手段对梅子70%乙醇部位进行分离纯化,从梅子提取物中分离纯化得到了14个化合物.运用核磁共振光谱、质谱等多种波谱方法,它们的化学结构分别鉴定为:5-羟甲基糠醛(1)、野黑樱苷(2)、顺式对羟基肉桂酸(3)、反式对羟基肉桂酸(4)、绿原酸(5)、柠檬酸(6)、柠檬酸单乙酯(7)、邻苯二酚(8)、原儿茶醛(9)、原儿茶酸(10)、新绿原酸(11)、咖啡酸(12)、山柰酚(13)、芹菜素(14).其中,化合物2~4及化合物8~9为首次从梅子中分离得到.从梅子中分离的所有化合物在合适的用量下均有良好的感官活性,可以提升卷烟的口感.

不同萃取方法下绿萼梅花蕾挥发性成分的差异研究

2023年3月13日,从浙江省长兴县东方梅园有限公司梅花基地采集绿萼梅花蕾,采用顶空-固相微萃取、水蒸气蒸馏、超临界二氧化碳3种萃取方法,结合气相色谱-质谱联用仪(GC-MS)分析绿萼梅Prunus mume f.viridicalyx花蕾中的挥发性成分,以明确其花蕾中挥发性成分组成、含量及不同萃取方式对挥发性成分萃取效果的影响。结果表明,三种萃取方法获得的绿萼梅花提取物中共鉴定出醛、醇、酮、烯等9类57种化合物,主要挥发性成分是苯甲醛、壬醛、金合欢醇、苯甲酸、α-香树脂醇、D-柠檬烯,其中苯甲醛含量最高。绿萼梅花顶空-固相微萃取物中共鉴定出21种化合物,以醛、醇和酯类化合物为主;水蒸气蒸馏物中共鉴定出30种化合物,以酯、烯、酮等水难溶化合物为主;而超临界二氧化碳提取物中共鉴定出30种化合物,以长链烷、酯类化合物为主。三种提取方法中超临界二氧化碳萃取物中蜡质含量较高,顶空-固相微萃取和水蒸气蒸馏法结合更适合作为植物芳香类挥发物的检测手段。

我国梅树上病毒及类病毒的检测和鉴定

目前,我国梅树上的病毒种类及发生情况仍不完全清楚。本研究从北京、武汉、南京和无锡的梅园中采集了64份疑似感染病毒的叶片样品,通过RT-PCR和斑点杂交,对7种病毒和2种类病毒进行了检测。共检测到6种病毒和1种类病毒。其中,李属坏死环斑病毒(prunus necrotic ringspot virus, PNRSV)和桃潜隐花叶类病毒(peach latent mosaic viroid, PLMVd)为我国梅树上的首次检出。PNRSV、亚洲李属病毒2号(Asian prunus virus 2, APV2)、桃叶痘伴随病毒(peach leaf pitting-associated virus, PLPaV)的检出率高于30%。综合考虑病毒的分布及检出率,PLPaV、APV2、PNRSV和李树皮坏死茎痘伴随病毒(plum bark necrosis stem pitting-associated virus, PBNSPaV)是武汉、南京和无锡梅树上的主要病毒。此外,通过克隆和测序,获得了PLMVd和梅树病毒A(mume virus A, MuVA)的基因组,PLPaV的RNA1组分和PNRSV外壳蛋白(CP)基因序列。序列比较分析显示,我国PLMVd梅分离物和PNRSV梅分离物与我国桃分离物亲缘关系最近,表明PLMVd和PNRSV可能在梅和桃树间交互侵染;我国MuVA梅分离物序列与日本梅分离物序列的相似性高达98.56%;PLPaV梅分离物与我国桃分离物之间序列变异较大。上述结果不仅进一步明确了我国梅树上的病毒及类病毒种类和分布情况,而且有助于深入了解它们的流行与传播。

不同梅品种花果特性分析与评价

【目的】梅Prunusmume是花果兼用型植物,在国内广泛应用。评价不同梅品种,为花果兼用梅品种选育以及梅果实加工利用提供依据。【方法】以梅28个品种为研究对象,测定其花部性状、结实量、果实品质指标,运用主成分分析与聚类分析法进行综合评价。【结果】28个梅品种花径为19.14-31.38 mm,花瓣数量为5-23枚,花色以白色品种占比最大。不同梅品种结实量差异较大,其中‘丽江照水’‘玉龙红翡’‘玉龙绯雪’易结实。不同梅品种果实外观品质与内在品质均存在差异。对28个品种进行主成分分析,16个指标简化为4个公因子,方差解释百分比为80.77%;综合得分表明:‘蝶羽重’‘骨里红’‘单粉垂枝’‘红颜朱砂’‘ZAFU-CZ02’‘江梅’‘丽江照水’得分较高;对28个品种进行系统聚类分析,结果表明:当欧氏距离为24时,梅品种可分为6类。综合主成分与聚类分析结果,筛选出‘单粉垂枝’‘丽江照水’‘骨里红’‘红颜朱砂’4个优良的花果兼用梅品种。【结论】不同梅品种花部性状、结实量及果实品质均存在差异。综合花部性状、结实量、果实品质可知:‘单粉垂枝’‘丽江照水’花单瓣,易结实,果实综合品质最佳,适宜作为果品加工的花果兼用梅品种;‘骨里红’‘红颜朱砂’花色艳丽,半重瓣,较易结实,适宜作为实生选种的花果兼用梅品种。