Research Papers

• Primary Analysis of the Function of bZIP Transcription Factor Gene MdAREB2 in Apple
• LIU Yajing，MA Qijun，LU Jing，HAO Yujin，and YOU Chunxiang*
• Acta Horticulturae Sinica. 2018, 45(1): 1-10. DOI:10.16420/j.issn.0513-353x.2017-0195
• Abstract ( 491 ) HTML ( 1167 ) PDF (2212KB) ( 1167 ) 　　　
• A bZIP transcription factor gene MdAREB2（MDP0000248567）was cloned from‘Gala’apple（Malus × domestica Borkh.）. The evolutionary tree analysis indicated that MdAREB2 had the very high homology with AtAREB2. The promoter of MdAREB2 was analyzed and predicted by PlantCare databases，and the results showed that the promoter sequence contained abscisic acid response element ABRE. Quantitative real-time PCR showed that MdAREB2 was significantly induced by ABA. This study found that with 0.5 μmol · L-1 ABA and 2 μmol · L-1 ABA treatment，excessive expression of MdAREB2 in mutant abi5 could restore its sensitivity to ABA in seed germination stage. In the seedling growth stage，with 20 μmol · L-1 ABA treatment，excessive expression of MdAREB2 in mutant abi5 could partially restore its sensitivity to ABA.
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• Dazomet Fumigation and Short-time Allium fistulosum Rotation Can Significantly Reduce the Apple Replant Disease
• XU Shaozhuo1，LIU Yusong1，XIA Mingxing2，WANG Yingping2，CHEN Xuesen1，SHEN Xiang1，YIN Chengmiao1,*，and MAO Zhiquan1,*
• Acta Horticulturae Sinica. 2018, 45(1): 11-20. DOI:10.16420/j.issn.0513-353x.2017-0176
• Abstract ( 268 ) HTML ( 867 ) PDF (895KB) ( 867 ) 　　　
• In order to provide basis for preventing and controlling apple replant disease，the Malus hupehensis Rehd.，which was planted in a Allium fistulosum rotation after dazomet fumigation under pot condition，was tested to study the effect on the soil environment and the growth of its seedling. Experiments were begun to conduct in May 2015，and four treatments were：soil of old apple orchard， Allium fistulosum rotation，dazomet fumigation，and Allium fistulosum rotation after dazomet fumigation. The seedlings of M. hupehensis Rehd. for all treatments were planted in May 2016. The results showed that，compared Allium fistulosum rotation after dazomet fumigation with the control in 2015，the fungi number was reduced by 73%；the copy number of Fusarium proliferatum which was analysised by Real-time PCR was reduced by 67%；the number of bacteria was reduced by 19%；the ratio of bacteria and fungi was increased by 213%. The results of principal component analysis of T-RFLP（PCA）and cluster analysis showed that，Allium fistulosum rotation after dazomet fumigation significantly changed soil fungal community structure. Compared with dazomet fumigation，the soil enzyme activity（urease，sucrase，catalase，phosphatase）of Allium fistulosum rotation after dazomet fumigation was increased. In August 2016，compared with the control，the plant height，stem diameter，fresh weight and dry weight of M. hupehensis Rehd. seedlings with a Allium fistulosum rotation after dazomet fumigation were 1.96，1.75，2.31 and 2.34 times those of the control，respectively，and the root length，root area，root volume，root respiration rate were increased by 155%，310%，294% and 89%，respectively. The treatment with a short time Allium fistulosum rotation after dazomet fumigation had the highest ratio of bacteria and fungi，significantly changed the soil fungal community structure，improved soil enzyme activity，promoted the growth of M. hupeheusis Rehd.，and had the better function in prevention and control of apple replant disease than that of dazomet fumigation，therefore，it could be an effective measure to reduce apple replant disease.
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• Phloridzin and Fusarium moniliforme Aggravated the Replanted Soil Environment and Inhibited the Growth of Malus hupehensis Seedlings
• JIANG Weitao1,*，YIN Chengmiao1,2,*，DUAN Yanan1，XIANG Li1，WANG Mei1，CHEN Xuesen1，SHEN Xiang1，ZHANG Min2,**，and MAO Zhiquan1,**
• Acta Horticulturae Sinica. 2018, 45(1): 21-29. DOI:10.16420/j.issn.0513-353x.2017-0285
• Abstract ( 301 ) HTML ( 804 ) PDF (721KB) ( 804 ) 　　　
• To provide a theoretical basis for clarifying the mechanism of apple replant disease，a pot experiment was conducted to study effects of phloridzin and Fusarium moniliforme on the growth of Malus hupehensis Rehd. seedlings and the replanted soil microorganisms. The determination results of the August showed that the biomass of M. hupehensis Rehd. seedlings was significantly reduced with the treatments of Fusarium moniliforme，phloridzin and Fusarium moniliforme + phloridzin. The plant height，ground diameter，fresh weight and dry weight of the Fusarium moniliforme + phloridzin were 70.6%，77.4%，52.3%，and 43.7% of the replanted soil control，respectively. The growth trend of the treatments in September was consistent with that in the year of August. Treatments that caused damage to the root respiration rate of M. hupehensis Rehd. seedlings in August and September were sorted as：Fusarium moniliforme + phloridzin > Fusarium moniliforme > phloridzin > replanted soil control. Three treatments inhibited the growth of bacteria in the replanted soil，promoted the growth of fungi and changed the community structure of soil fungi. The activities of soil catalase，soil urease，soil invertase，soil neutral phosphatase were significantly reduced with the three treatments；In August，soil urease was reduced by 14.8%，29.1%，53.4% compared with the replanted soil control. In conclusion，Fusarium moniliforme and phloridzin alone or in combination significantly inhibited the growth of M. hupehensis Rehd. seedlings，Fusarium moniliforme and phloridzin treatment was the most significant. These results suggested that the combination of phloridzin and Fusarium moniliforme could increase the phenomenon of apple replant disease.
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• Effects of Salt Stress on the Generation and Scavenging of Reactive Oxygen Species in Leaves of Grape Strains with Different Salt Tolerance
• FU Qingqing，TAN Yazhong，ZHAI Heng，and DU Yuanpeng*
• Acta Horticulturae Sinica. 2018, 45(1): 30-40. DOI:10.16420/j.issn.0513-353x.2017-0388
• Abstract ( 423 ) HTML ( 1127 ) PDF (4242KB) ( 1127 ) 　　　
• In order to study the generation and scavenging of reactive oxygen species（ROS）in leaves of grape strains with different salt tolerance under salt stress，the salt-tolerance tissue culture hybrids（A15 and A17）of Vitis amurensis Rupr.‘Zuoshan 1’× SO4 were used as materials to be irrigated with 100 mmol · L-1 NaCl，and tissue culture 1103P was used as negative control. O2- production rate，H2O2 content，MDA content，relative electrical conductivity，chlorophyll content，antioxidant enzyme activities and antioxidant substances were determined. The results showed that，under salt stress，SOD，CAT and POD activities，AsA and GSH content and APX and GR activities in leaves were higher in salt-tolerance hybrids A15 and A17，ROS and MDA content were lower，and the increment of relative electrical conductivity was less，the structure of chloroplast was integrated，and the chloroplast content was higher；while the overproduction of ROS and MDA damaged the membrane integrity and reduced chloroplast content in 1103P. Salt-tolerance hybrids A15 and A17 remained high antioxidase activity and AsA-GSH cycle efficiency，reduced the damage of membrane caused by lipid peroxidation，maintained the structure and function integrity of chloroplast.
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• Studying Function of Alcohol Acyltransferase Gene FvAATW2 of Fragaria vesca by Over-expressing in Tobacco and Cultivated Strawberry
• DONG Jing，WANG Guixia，ZHONG Chuanfei，CHANG Linlin，SUN Jian，ZHANG Hongli，SUN Rui，SHI Kun，WEI Yongqing，and ZHANG Yuntao*
• Acta Horticulturae Sinica. 2018, 45(1): 41-50. DOI:10.16420/j.issn.0513-353x.2017-0085
• Abstract ( 281 ) HTML ( 862 ) PDF (1095KB) ( 862 ) 　　　
• FvAATW2 was an alcohol acyltransferase gene，cloned from Fragaria vesca L. fruits by RT-PCR. In this study，the sense plant expression vector pBI121-FvAATW2 driven by cauliflower mosaic virus 35S promoter was constructed through replacing GUS with FvAATW2，and then was used to genetically transform tobacco and a cultivated strawberry cultivar‘Camarosa’，to study the function of FvAATW2. Agrobacterium tumefaciens strain LBA4404 carrying pBI121-FvAATW2 was employed to infect leaf-discs in the genetic transformation. Transgenic lines were identified by both PCR and Southern blot. The expression of exogenous FvAATW2 in transgenic in vitro plants was studied using relative enzyme activity and real-time quantitative PCR. It was found that FvAATW2 expressed in all 6 tested tobacco lines and AAT activity values of 5 lines were higher than wild-type plants. Volatiles in leaves of the transgenic tobacco with the highest AAT activity were also detected by SPME/GC-MS at 10 ~ 12 leaves phase. Octyl propanoate was found in transgenic leaves，while not any ester captured in wild-type plants at the same phase. The results showed that pBI121-FvAATW2 functioned correctly and esters could be synthesized much earlier in transgenic tobacco than in wild-type one. Four strawberry transgenic lines were obtained after PCR and Southern blot operation. A transgenic line bearing more aromatic fruits was screened out and used for the volatiles detection. Through analyzing the ester composition of mature transgenic fruits，it was found that the proportion of esters in total volatiles and respective proportions of octyl acetate，ethyl hexanoate，octyl hexanoate，and ethyl octanoate were higher significantly than those of wild-type fruits，while mass fraction of methyl butanoate decreased significantly and that of ethyl octanoate increased significantly. It was indicated that exogenous FvAATW2 expressed normally in cultivated strawberry and effected ester synthesis，and it could make fruit aroma intense by changing ester composition.
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• Effects of Energy Charge Levels and Expression of Energy Charge Genes on Browning of Post-harvest Litchi Fruit
• GAO Zhaoyin1，HU Meijiao1，ZHAO Chao2，ZHANG Zhengke3，LI Min1，LI Huanling1，WANG Guo1，SUN Jinhua1，WEN Jie2，and WANG Jiabao1,*
• Acta Horticulturae Sinica. 2018, 45(1): 51-60. DOI:10.16420/j.issn.0513-353x.2016-0930
• Abstract ( 286 ) HTML ( 873 ) PDF (890KB) ( 873 ) 　　　
• To discover the mode of energy regulation and its relationship with senescence in harvested Litchi chinensis Sonn.，three energy charge（EC）levels of seedless litchi fruit including normal EC at ambient temperature（25 ℃）storage（the control），lower EC as affected by 2,4-Dinitrophenol（DNP）and higher EC at low temperature（5 ℃）were investigated to determine effects of EC levels and energy-related genes expression on pericarp browning and senescence. The results showed that low EC level under DNP treatment remarkably promoted fruit senescene；accelerated senescence was observed in the control fruits during post-storage，which was associated with decreased EC；and delayed senescence was noted in 5 ℃-stored litchi fruit with higher EC. DNP treatment resulted in up-regulated expression of LcAOX1 within 6 h，and severe browning in DNP-treated fruit initiated after 24 h of storage. DNP treatment strongly inhibited the expression of LcUCP1. Compared to the control fruits，DNP treatment resulted in earlier and more intense expression in LcSnRK2. During post-storage，lower EC levels were observed in both control and DNP-treated fruits，and expressive levels of LcAAC1，LcAOX1 and LcUCP1 appear to decrease while fruit senescence was aggravated. The expressions of LcAAC1，LcAOX1，LcUCP1 and LcSnRK2 in fruit stored at 5 ℃ were maintained at relatively low levels throughout the storage. The present results indicate that the decline of EC level in harvested litchi fruit might be one of the most important factors to cause rapid senescence. The genes including LcSnRK2 and LcAAC1 were sensitive to energy changes in litchi pericarp，which might respond rapidly to deficit of the energy，and might also be induced to express in up-regulation. The up-regulation of LcAOX1 and LcUCP1 was positively correlated with the senescence of litchi fruit，which might be as reference indicators for postharvest senescence of litchi fruit.
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• Comparison of Seed-setting and the Transmission Rate of Rfo During the Distant Hybridization of Chinese Kale × Brassica napus and Cabbage × B. napus
• YU Hailong*，LI Zhiyuan*，YANG Limei，LIU Yumei，ZHUANG Mu，Lü Honghao，LI Zhansheng，FANG Zhiyuan**，and ZHANG Yangyong**
• Acta Horticulturae Sinica. 2018, 45(1): 61-70. DOI:10.16420/j.issn.0513-353x.2017-0557
• Abstract ( 677 ) HTML ( 995 ) PDF (4432KB) ( 995 ) 　　　
• In order to determine whether the seed-setting of distant hybridization between Brassica napus and B. oleracea is affected by different B. oleracea sub-species or not，in this study，two Chinese kale and four cabbage materials with Ogura CMS cytoplasm were crossed with two homozygous rapeseed restored lines harboring the Ogura CMS fertility-restored gene Rfo. The F1 interspecific hybrid plants with doubled chromosome were backcrossed with Ogura CMS Chinese kale and cabbage to produce BC1 plants，respectively. The seed-setting and the transmission rate of Rfo of Chinese kale × rapeseed cross combinations（CKR）and cabbage × rapeseed cross combinations（CR）were investigated in two generations，respectively. The results suggested that the compatibility index of CKR was two folds higher than that of CR，with significant difference between CKR and CR in two generations（P < 0.05）. In addition，the Rfo gene transmission rate of CKR was higher than that of CR. The pollen viability and recovering ratio of genetic background（RRGB）of all BC1 Rfo-positive individuals obtained from CKR and CR were assessed. The pollen viability of most BC1 Rfo-positive individuals obtained from CKR were significantly higher than that of the two Rfo-positive individuals obtained from CR（P < 0.05）. The RRGB of 26 Rfo-positive individuals obtained from CKR were 0.50–0.84. While the RRGB of two Rfo-positive individuals（15Y1 and 15Y4）obtained from CR were 0.53 and 0.59，respectively. Our results revealed that compared with cabbage，Chinese kale is much easier to produce progenies when crossed with B. napus and the transmission rate of alien fragment was much higher. Therefore，Chinese kale could be used as a bridge to accelerate the process of the beneficial agronomic traits transfer from B. napus to cabbage.
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• Transcriptome Analysis of Stigma After Pollination in Brassica oleracea var. capitata
• PU Min1，LIAN Xiaoping2,*，LUO Shaolan1，ZHANG Hecui1，WANG Yukui1，BAI Xiaojing1，ZENG Jing1，GAO Qiguo2，REN Xuesong2，and ZHU Liquan1,**
• Acta Horticulturae Sinica. 2018, 45(1): 71-84. DOI:10.16420/j.issn.0513-353x.2017-0480
• Abstract ( 436 ) HTML ( 1617 ) PDF (803KB) ( 1617 ) 　　　
• Illumina HiSeq2000，a high-through transcriptome sequencing technology，was applied to obtain the transcriptome differential expression data of stigma after self-pollinated 30 min，cross-pollinated 30 min and unpollinated. The objective of this study is to analyze the differences of gene expression of stigma after self-pollination and cross-pollination in Brassica oleracea L. var. capitata，and further analyze the significantly different expression genes. Transcriptome sequencing obtains a total of 2.38 × 1010 bp bases containing 1.6 × 108 of the original pair-end reads. There were 2 900 and 2 328 genes with altered expression in stigma after self-pollination and cross-pollination in Brassica oleracea L. var. capitata，respectively，and a total of 1 904 common genes expressed between two samples. The greater proportional difference after self-pollination is cell killing，extracellular matrix part and nucleic acid binding transcription factor activity. The greater proportional difference after cross-pollination is the signaling，extracellular region part，nutrient reservoir activity and structural molecule activity in the background of differential expression genes and all genes. GO enrichment analysis illustrated that the 40 specific highly significant GO terms in self-pollination sample mainly involved in sugar transmembrane transporter activity，tubulin binding and calmodulin-dependent protein kinase activity. The 53 specific highly significant GO terms in cross-pollination sample mainly involved in farnesoic acid O-methyltransferase activity，minus-end-directed microtubule motor activity and auxin influx transmembrane transporter activity. About 563 and 453 genes their expression was significantly up-regulated and down-regulated respectively in the 996 specific differentially expressed genes in self-pollination sample，which mainly related to calciumion binding，cytoskeletal correlation，jasmonic acid mediated signalin0g pathway and salicylic acid mediated signaling pathway. The 424 specific differentially expressed genes in cross-pollination sample mainly involved in transmembrane transport of substance and lipid metabolism．
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• Molecular Cloning，Location and Expression Analysis of BoPLL in Self-incompatibility Brasscia oleracea
• LUO Shaolan1，PU Min1， ZENG Jing1，SHI Songmei2， LIAN Xiaoping2，WANG Yukui1，BAI Xiaojing1，ZHANG Hecui1，and ZHU Liquan1,*
• Acta Horticulturae Sinica. 2018, 45(1): 85-96. DOI:10.16420/j.issn.0513-353x.2017-0389
• Abstract ( 319 ) HTML ( 889 ) PDF (2239KB) ( 889 ) 　　　
• The stigmas were from collected self-pollination and cross-pollination of Brassica oleracea L. var. capitata L. and were used to analyze protein expression profiles. We obtained a protein that was down-regulation expression in self-pollination and up-regulation in cross-pollination treatments，which was named BoPLL. Moreover，transcriptomic analysis also showed BoPLL was up-regulated at 0–30 min and down-regulated at 30–60 min in self-pollination process，while it was up-regulated in cross-pollination process. The CDS of BoPLL was 1 212 bp in length，encoding 403 amino acids of BoPLL. Sequence analysis found BoPLL contained a high conserved PASTA domain and a CMM-10 domain，moderate conserved PbH1 domain，and a low conserved HYDRO domain，indicating it was a typical PLL protein. Chromosomal location displayed that BoPLL is not linked with S locus genes. Phylogenetic tree analysis showed that Brassica oleracea L. BoPLL was more close to Arabidopsis thaliana AtPLL rather than Medicago truncatula MtPLL. RT-PCR analysis found BoPLL was predominantly expressed in pollen and stigma，with lower expression level of BoPLL in pistil than pollen，and no expression in leaf，flower bud，sepal and petal. qRT-PCR analysis revealed that the mRNA in 15~60 min expression patterns of BoPLL was similar with the RNAseq analysis. Based on those results，we speculated that BoPLL was involved in SI response in B. oleracea.
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• Studies on Identification Methods in Backcross Generation of Cry1Ac-transgenic Broccoli
• LI Zhansheng，ZHANG Lili，ZHANG Xiaoli，SHU Jinshuai，SU Yanbin，SUN Jifeng，LIU Yumei*，FANG Zhiyuan，YANG Limei，ZHUANG Mu，ZHANG Yangyong，and Lü Honghao
• Acta Horticulturae Sinica. 2018, 45(1): 97-108. DOI:10.16420/j.issn.0513-353x.2017-0617
• Abstract ( 328 ) HTML ( 747 ) PDF (1954KB) ( 747 ) 　　　
• It’s important for separation offspring in transgenic Bt gene broccoli to identify positive plant lines and resistant materials accurately by different methods. In the study，five backcross materials based on four to five generations，one homozygous material as positive control and five inbred lines as negative controls were used for analysis of the insect-resistant effectiveness of Cry1Ac in transgenic broccoli from the levels of gene，protein and appearance. The gene of Cry1Ac was carried out by PCR amplification，protein was carried out by determination of Bt-Cry1Ab/1Ac test strip，and appearance was carried out by feeding test of diamondback moth in vitro. At the same time，the advantages of three methods were evaluated and stated. The result showed that a total of 82，77 and 75 positive strains were divided from 159 strains of five backcross materials. According to chi-square test at the level of 5%，the positive and none positive strains of 1︰1 were calculated and in accordance with mendelian inheritance. By comparisons of three methods，we found that the feeding trial of diamondback moth in vitro was accurate and reliable，but the experimental conditions must to be well controlled. The Bt-Cry1Ab/1Ac test strip was sensitive，fast and can observe the actual Bt protein expression，but it must control human errors. The normal PCR amplification could be used to identify the positive strains，but it can not reflect the actual anti-insect effect，so it needs to be together with the test of feeding diamondback moth or the test of Bt protein，which could provide an accurate result.
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• Ethylene Responsive Factors ERF Regulated the Hypoxia Response of Transformed Chrysanthemum Lines
• LIU Xiaofen，XIANG Lili，YIN Xueren，LI Fang，and CHEN Kunsong*
• Acta Horticulturae Sinica. 2018, 45(1): 109-116. DOI:10.16420/j.issn.0513-353x.2017-0184
• Abstract ( 303 ) HTML ( 955 ) PDF (2734KB) ( 955 ) 　　　
• The exogenous DkERF9，a reported hypoxia responsive gene from persimmon，was transformed into‘Jiuyuejiu’chrysanthemum. Five positive lines（35S::DkERF9）were obtained and verified with PCR. Compared to the wild type plants，the growth of 35S::DkERF9 lines were inhibited significantly by water planting treatments for two weeks，as the length of roots and shoots were shorter in 35S::DkERF9 lines. The differences between 35S::DkERF9 lines and wild type plants remain significant，after three weeks recovery.
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• The Changes of Anthocyanin Content During the Development of Primulina swinglei Flower
• HU Bin1,2，DING Dehui1,3，FU Xiumin1，FENG Chao1,*，and KANG Ming1
• Acta Horticulturae Sinica. 2018, 45(1): 117-125. DOI:10.16420/j.issn.0513-353x.2017-0205
• Abstract ( 233 ) HTML ( 819 ) PDF (2203KB) ( 819 ) 　　　
• In this paper，visualization of tissue section and UPLC-Q-TOF-MS technology were applied to investigate differences in the overall anthocyanin content，distribution and composition in different organs and tissues during the development of Primulina swinglei flowers. Seventeen anthocyanins were detected and identified，including cyanidin，peonidin，delphinidin，petunidin and malvidin，which were found in their glycosylated forms linked with sugars such as glucoside，rutinoside，acetyl rutinoside glycosides and other aglycones. The purple color in petals is due mainly to the high proportion（up to 64.0%）of bluish violet pigment of delphinidin，petunidin and malvidin. While the red cylinder color is possibly due to cyanidin and peonidin，which accounted for 36.7% and 20.0%，respectively，of the total anthocyanins. Before blooming，the total anthocyanin increased slowly and then became stable. The percentage of the red pigmented compounds decreased and bluish violet pigments increased during flower development，probably due to the increased contribution of petals to the total mass of the flower. After blooming，the content of anthocyanin decreased，probably due to the pigment degradation or dilution.
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Research Notes

• Genetic Diversity and Relationship Analysis Among Cerasus pseudocerasus，C. avium，and C. tomentosa Based on Internal Transcribed Spacer（ITS）Sequences
• WANG Hao1，HUANG Zhilin1，CHEN Tao2，ZHANG Jing2，WANG Yan2，CHEN Qing1，TANG Haoru1,2，and WANG Xiaorong1,2,*
• Acta Horticulturae Sinica. 2018, 45(1): 126-138. DOI:10.16420/j.issn.0513-353x.2017-0317
• Abstract ( 437 ) HTML ( 1056 ) PDF (1535KB) ( 1056 ) 　　　

• In this study，we sampled a total of 95 samples，including 35 landraces and 35 wild individuals of Chinese cherry from 6 populations across 35 counties of 8 provinces in China，18 cultivars of European sweet cherry from 10 countries in the world，as well as 7 semi-wild Nanking cherry resources from 7 counties of 5 provinces in China. Based on internal transcribed spacer（ITS）sequences，we tried to study the genetic diversity within species and interspecific relationship among them. The final ITS dataset contained 712 aligned nucleotides，with 58.1% of G + C content，of which 71（9.97%）were variable sites. The ITS sequences defined 37 haplotypes. The haplotype diversity（Hd）and nucleic acid diversity（π）of C. pseudocerasus，C. avium，and C. tomentosa were（0.840，0.00466），（0.928，0.00396）and（0.905，0.00564），respectively. Within C. pseudocerasus，the genetic diversity of germplasm（Hd = 0.726，π = 0.00224）was significantly lower than that of wild resource（Hd = 0.914，π = 0.00681）. Interspecific genetic relationship analysis showed close relationship（0.019）between Chinese cherry and sweet cherry，while they revealed further relationship with Nanking cherry. Both Neighbor-Joining reconstruction and haplotype network showed three distinct branches with obvious genetic differentiation among the three species. In addition，there were significant differences in the secondary structure of the ITS1 and ITS2 regions and t-detection of the minimum free energy. Based on above results，there were high genetic diversity and obvious genetic differentiation among three cherry cultivated species. Close relationship was detected between Chinese cherry and European sweet cherry，while far distance between Nanking cherry with them.

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• Analysis of SSR Markers Information and Primer Selection from Transcriptome Sequence of Hybrid Hazelnut Corylus heterophylla × C. avellana
• CHENG Yunqing，ZHANG Lina，ZHAO Yongbin，and LIU Jianfeng*
• Acta Horticulturae Sinica. 2018, 45(1): 139-148. DOI:10.16420/j.issn.0513-353x.2017-0281
• Abstract ( 298 ) HTML ( 831 ) PDF (690KB) ( 831 ) 　　　
• Deep bio-information analysis was carried out using transcriptome data in order to develop SSR marks in hybrid hazelnut Corylus heterophylla × C. avellana. In total，35 604 SSR sequences were found in 25 649 SSR containing Unigenes. In all the SSR sequences，mono-nucleotide，dinucleotide and trinucleotide repeats belonged to high frequent type，and their number and percent were 10 102（28.37%），16 543（46.46%）and 7 035（19.76%）respectively. In mono-nucleotide repeats，A/T was most abundant，accounting for 28.13% of the total SSR amount. In dinucleotide repeats，AG/CT，AT/AT，AC/GT were more abundant，accounting for 35.79%，7.47% and 3.17% of the total. In repeats of trinucleotide，tetranucleotide，pentanucleotide and hexanucleotide，the highest percent sequence of each nucleotide repeat were AAG/CTT，AAAG/CTTT，AAAAG/CTTTT，AAAAAG/CTTTTT，and they accounted for 7.24%，0.33%，0.38% and 0.15% of the total SSR amount respectively. Sixty pairs of randomly chosen SSR primers were synthesized and amplified from 25 Corylus samples，including Corylus heterophylla Fisch，C. heterophylla × C. avellana，C. avellana and C. mandshurica Maxim. Polymorphism information content（PIC）values of seven polymorphic pairs of primers varied from 0.35 to 0.70，and 52 polymorphic bands were obtained. UPGMA（Unweighted Pair Group Method Analysis）clustering results indicated that these primers could precisely separate germplasms mentioned above into different groups. This study clarifies SSR distribution characteristics of hybrid hazelnut C. heterophylla × C. avellana and provides a scientific basis for further studies of genetic linkage map construction in these species.
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• Excavation and Usablility Analysis of SSR from Transcriptome of Clausena lansium
• LU Yusheng，CHEN Zhe，CHANG Xiaoxiao，QIU Jishui，LIN Zhixiong*，and PAN Jianping*
• Acta Horticulturae Sinica. 2018, 45(1): 149-158. DOI:10.16420/j.issn.0513-353x.2017-0342
• Abstract ( 253 ) HTML ( 778 ) PDF (1240KB) ( 778 ) 　　　
• In this study，a total of 68 998 Unigenes from transcriptome of Clausena lansium were used for screening of SSR loci using MISA software. 11 825 SSRs，including the types of 1–6 nucleotide repeats distributed in 11 000 Unigenes were identified，with occurring frequency of 17.14% and mean distance of 4.41 kb. The major types of SSR loci were 1–3 nucleotide repeats，accounting for 93.93% of the SSR loci，of which mono-，di- and tri- nucleotide repeats were 41.48%，24.92% and 27.53%，respectively. Two hundred and seven repeat motifs with iteration numbers from 4 to 24 were discovered from the 11 825 SSR loci，and the most abundant motif were A/T，AG/CT，AT/AT，AAG/CTT and AAT/ATT. The lengths of repeat nucleotide sequences for all SSR loci ranged from 12 to 25 bp，with most from 12 to 15 bp，accounting for 50.95% of all SSR loci. A total of 6 102 SSR primers were designed and 30 primers were selected randomly for PCR amplification tests，of which 24 primers amplified effective and 13 primers showed polymorphism in sixteen different wampee germplasm. The results indicated that the Unigenes generated from trasncriptome sequencing in C. lansium could be used as an effective source to develop SSR markers. The large quantities of SSR markers will provide reliable markers for map structure，genetic polymorphism analysis for wampee germplasm resource.
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• Development of SSR Molecular Markers Based on Transcriptome Sequencing of Tagetes erecta
• ZHANG Huali1，CONG Richen1，WANG Maoliang1，Dong Aixiang1，XIN Haibo1,*，YI Mingfang2，and GUO Hua3,*
• Acta Horticulturae Sinica. 2018, 45(1): 159-167. DOI:10.16420/j.issn.0513-353x.2017-0166
• Abstract ( 399 ) HTML ( 860 ) PDF (1483KB) ( 860 ) 　　　
• A total of 48 953 Unigenes were obtained by transcriptome sequencing of flower buds of Tagetes erecata. MISA found 20 666 SSRs located in 13 849 unigenes with the frequency of 28.29% and a mean distance of 2.51 kb per one. Trinucleotide and tetranucleotide were major types with the percentages of 50.16% and 20.94%，respectively. ATG/ATG and AAAC/GTTT were most frequent motifs in trinucleotide and tetranucleotide repeats，accounting for 13.82% and 3.66%，respectively. Based on different dominant motifs types，36 primers were chosen randomly and verified with 20 different inbred lines of Tagetes erecta. The results showed that 30 primers were effective with a valid rate of 80.56%，and 13 primers showed polymorphism with He and PIC mean values of 0.275 and 0.608，respectively. These data indicated that the Unigenes generated from transcriptome sequencing can be used as an effective source to develop SSR markers，and the large quantities of SSR markers will provide reliable resources for analysis on genetic polymorphism and constructing genetic map in Tagetes erecta.
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• Establishing Virus Induced Gene Silencing（VIGS）System in Tree Peony Using PsUFGT Genes
• SHU Qingyan1,*，ZHU Jin1,2,*，MEN Siqi1,2，HAO Qing3，WANG Qianyu1,2，LIU Zheng’an1，ZENG Xiuli4，and WANG Liangsheng1,2,**
• Acta Horticulturae Sinica. 2018, 45(1): 168-176. DOI:10.16420/j.issn.0513-353x.2017-0522
• Abstract ( 338 ) HTML ( 1028 ) PDF (734KB) ( 1028 ) 　　　
• Considerable progress has been made on the phytochemical basis of flower color formation on tree peony（Paeonia suffruticosa Andrews）. However，due to lack of genetic transformation system，function of the genes related to flower color formation has to be characterized using other model plant systems. We utilized virus induced gene silencing（VIGS）for establishing the functional characterizing technique to study UDP-glucose：flavonoid glycosyltransferases（UFGTs）genes in tree peony. The fragments with 413 bp and 418 bp length with a putatively conserved domain of PsUF3GT and PsUF5GT were selected for constructing VIGS recombinant vectors，respectively. Nine treatments with two factors and three levels were conducted to test the gene silencing effects. The results showed that the expression level of PsUF3GT after infiltrating petals of blooming for 15 min，or the expression level of PsUF5GT after infiltrating petals of bud stage for 10 min，were reduced 65.0% and 85.0% as compared with that of control petals of blooming or bud stage infiltrated using empty vectors for 10 min or 15 min，respectively. Meanwhile，total anthocyanins content in petal blotch were also decreased 24.2% and 28.2% after treatment of flowers at blooming or bud stage by infiltration for 15 min or 10 min，respectively，in which，cyanidin 3-O-glucoside reduced 92.2% in petals at blooming stage infiltrated for 15 min and cyanidin 3,5-O-diglucoside reduced 54.9 % in petals at bud staged infiltrated for 10 min. Therefore，an optimal VIGS system has been established in tree peony，namely，flowers at blooming or bud stage were subjected to vacuum infiltration for 10–15 min，then kept in dark for 1 d at 8 ℃ with relative 60% humidity，afterwards，kept at 23–25 ℃ with normal light and relative 60% humidity for 3 d，then，treated samples could be used for further functional characterization of the silenced genes. Our VIGS method would be beneficial to functional characterization of the genes related to flower color formation，and study the molecular mechanism of flower color variation.
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Review

• Biosynthesis of Flavonol and Its Regulation in Plants
• CAO Yunlin，XING Mengyun，XU Changjie，and LI Xian*
• Acta Horticulturae Sinica. 2018, 45(1): 177-192. DOI:10.16420/j.issn.0513-353x.2017-0306
• Abstract ( 462 ) HTML ( 1507 ) PDF (973KB) ( 1507 ) 　　　
• Flavonol has a wide range of biological activities and studies on biosynthesis and regulation of flavonol in plant attract more attention. Here，research progress in key enzymes（FLS，F3H，F3′H，F3′5′H，UGT）and regulatory factors（e.g. MYB transcription factors）in flavonol biosynthetic pathway were summarized from biochemical and molecular biological points of view. Important evidence illustrating key structure genes or regulatory factors playing critical roles in flavonol biosynthesis are emphasized，which may result in efficient regulation of target molecules and accumulation of flavonol compounds with important biological functions via genetic engineering.
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New Cultivars

• A New Early-maturing Red-flesh Peach Cultivar‘Zaoxianhong’
• WANG Furong，GONG Linzhong，WANG Huiliang，LIU Yong，AI Xiaoyan，GU Xia，LIU Mofa，and HE Huaping*
• Acta Horticulturae Sinica. 2018, 45(1): 193-194. DOI:10.16420/j.issn.0513-353x.2017-0015
• Abstract ( 403 ) HTML ( 691 ) PDF (1290KB) ( 691 ) 　　　
• ‘Zaoxianhong’is a new early-maturing red-flesh peach cultivar which is derived the cross of‘Shuguang’and‘Red-flesh Peach 18’. The fruit is round in shape，weighting 103 g on average，reaching up to 130 g. The flesh is red，crisp，sweet，with 10%–11% soluble solids content and 0.15% titratable acid content. The stone is cling. This cultivar appeared high and steady yield. The average yield is 22.5 t · hm-2.
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• A New Carya cathayensis Cultivar‘Yayou 8’
• CHANG Jun1，YAO Xiaohua1，LANG Xuejun2，WANG Kailiang1，TENG Jianhua3，WANG Nianjin4，and REN Huadong1,*
• Acta Horticulturae Sinica. 2018, 45(1): 195-196. DOI:10.16420/j.issn.0513-353x.2017-0160
• Abstract ( 297 ) HTML ( 654 ) PDF (1091KB) ( 654 ) 　　　
• ‘Yayou 8’is a new cultivar of Carya cathayensis which was selected from wild resources of Carya cathayensis in Jiande，Zhejiang Province. Compared to other cultivar of Carya cathayensis，the fruit of‘Yayou 8’was larger. The average weight was 5.72 g. The average fruit height was 25.19 mm. The average width of nut was 22.25 mm. And the fruit shape index was 1.13. The kernel accounted for 44.8% in whole fruit of‘Yayou 8’. The fat content of nutlets was 61.10%；the protein content was 65.30 mg · g-1；the potassium content was 2 820.2 μg · g-1，the calcium content was 1 275.7 μg · g-1. The average single fruit production of seven-year-old trees of‘Yayou 8’was 1 291.65 kg · hm-2. In a word，‘Yayou 8’has the good properties of early bearing，high yield，and excellent comprehensive characteristics.
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• A New Ganoderma lucidum Cultivar‘Panzhi 1’
• ZHOU Jie1，ZHANG Bo1，YANG Mei2，LIU Chengyi2，TAN Wei1，and LI Xiaolin1,*
• Acta Horticulturae Sinica. 2018, 45(1): 197-198. DOI:10.16420/j.issn.0513-353x.2017-0142
• Abstract ( 301 ) HTML ( 706 ) PDF (1194KB) ( 706 ) 　　　
• The new Ganoderma lucidum cultivar‘Panzhi 1’was derived from wild species collected from Fagaceae woods in Yanbian County of Panzhihua City，Sichuan Province. The pileus of‘Panzhi 1’was in kidney shape，yellow brown to red brown and glossy. The optimum growth temperature of mycelia and fructification stage was 24–26 ℃. The species could be cultured by bags and woods and field cultivation of‘Panzhi 1’was better. The average biological conversion rate of cultivation in woods and bags was 20%，29%，respectively.
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• A New Dendrobium Cultivar‘Jinyun’
• Acta Horticulturae Sinica. 2018, 45(1): 199-200. DOI:10.16420/j.issn.0513-353x.2017-0331
• Abstract ( 294 ) HTML ( 849 ) PDF (886KB) ( 849 ) 　　　
• The new cultivar‘Jinyun’is derived from the cross Dendrobium‘Yuki’and Dendrobium ‘Thai Red’. The flower color is purple and bright and the average single branch has 7.2 flowers with flower diameter of 7.0 cm. The beginning flower of this cultivar is in September and full bloom is in October and November when it cultured in greenhouse of Guangzhou. It can be widely planted in South China.
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