Research Papers

• Effects of Low Temperature Storage on Energy Metabolism，Related Physiology and Quality in‘Jinhong’Apple Fruit
• WANG Zhihua, JIA Chaoshuang, WANG Wenhui, TONG Wei, and JIANG Yunbin
• Acta Horticulturae Sinica. 2020, 47(12): 2277-2289. DOI:10.16420/j.issn.0513-353x.2020-0418
• Abstract ( 726 ) HTML ( 612 ) PDF (825KB) ( 612 ) 　　　
• In order to investigate the effect of low temperature storage on energy metabolism and physiological quality of‘Jinhong’apple fruit，‘Jinhong’apple fruit were stored at low temperature conditions of–2，0，2 and 4 ℃，and the energy-related substance was regularly measured such as adenosine triphosphate（ATP），adenosine diphosphate（ADP），adenosine monophosphate（AMP）content，energy charge（EC）and H+-ATPase，Ca2+-ATPase, succinate dehydrogenase（SDH）and cytochrome oxidase （CCO），superoxide dismutase（SOD）and other related enzyme activities. At the same time we measured the fruit respiration intensity，ethylene release，fruit hardness，vitamin C，titratable acid，soluble solids content and other physiological quality indicators，and investigated the fruit tissue browning index. The results showed that the ATP，ADP and AXP contents，EC and energy-related metabolic enzyme except Ca2+-ATPase activities，SOD activity and ethylene release of fruits stored at–2 ℃ were kept to the lowest throughout storage and shelf life，and the vitamin C content decreased rapidly in the later period. The fruit hardness，soluble solids content and titratable acid content remained the lowest at 4 ℃.The browning index of fruit at–2 ℃ was the highest（both skin and flesh were severely browned），the skin and flesh of the fruit stored at 0 ℃ had slight browning，and the fruit stored at 2 and 4 ℃ had no browning. During storage of 30，45，90 d，90 d + 3 d shelf life and 90 d + 5 d shelf life，fruit stored at 2 ℃ maintained higher energy levels，and fruit quality and flavor remained well，followed by fruit at 0 ℃. The results showed that there was a close relationship between tissue browning and energy deficit. The more energy deficit，the more serious browning. Appropriate low temperature can maintain a higher energy charge level of the fruit，while inhibiting pericarp browning，maintaining fruit storage quality，and delaying fruit senescence.
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• DRL1，a NAC Gene from Vitis vinifera‘Yatomo Rose’，Negatively Regulates the Drought Tolerance
• ZHU Ziguo, YIN Qizhong, ZHANG Qingtian, HAN Zhen, ZHANG Qian, and LI Bo,
• Acta Horticulturae Sinica. 2020, 47(12): 2290-2300. DOI:10.16420/j.issn.0513-353x.2020-0185
• Abstract ( 325 ) HTML ( 329 ) PDF (3553KB) ( 329 ) 　　　
• The function of NAC transcription factor DRL1，isolated from Vitis vinifera‘Yatomo Rose’，was characterized using the fluorescence quantitative PCR and transgenic technology. Under the hormone and stress treatments，the expression of DRL1 decreased with the strongest reduction in ABA and drought treatments. Exposed to ABA，DRL1 transgenic plants reduced the germination rate and root length. Treated with drought stress，DRL1 enhanced the sensitivity to drought tolerance and the stress related genes，such as NtLEA5，NtP5CR1，NtPSCS1，NtERD10C and NtDREB3，were down-regulated in transgenic plants. Moreover，DRL1 restrained the development of stele，and the vessel area was only 58% of wild tobacco. According to these results，DRL1 might negatively regulate drought stress.
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• Construction of Yeast Two-hybrid Three-frame cDNA Library of Vitis amurensis and Screening of VaCIPK18 Interaction Protein
• ZHENG Qiaoling, SHEN Wei, YAO Wenkong, and XU Weirong,
• Acta Horticulturae Sinica. 2020, 47(12): 2301-2316. DOI:10.16420/j.issn.0513-353x.2020-0115
• Abstract ( 398 ) HTML ( 456 ) PDF (4222KB) ( 456 ) 　　　
• The CBL-CIPK signaling system is comprised of calcineurin B-like protein（CBL）and its interacting protein kinases（CIPK），playing critical roles in response to calcium signal transduction and various stress responses in eukaryotes. A yeast two-hybrid（Y2H）three-frame cDNA library with the leaves of Vitis amurensis Rupr.‘Shuangfeng’under low temperature stress was constructed using SMART and LD-PCR techniques. The capacities of three-frame libraries were 1.7 × 106，1.3 × 106 and 1.9 × 106 cfu • mL-1，respectively，with the inserted fragment of 500–2 000 bp in length，and 100% recombination rate，which could meet with the requirements of yeast two-hybrid screening. A low-temperature-induced gene VaCIPK18 that previously identified by our team，which localized at both nucleus and cytoplasm，was used as a“bait”to obtain three constructs containing the full length cDNA，as well as the two truncations of the S_TKc domain（VaCIPK18△S_TKc）and the NAF domain（VaCIPK18△NAF），respectively. The result showed that a total of 17 candidate interacting proteins of VaCIPK18 were screened from the library，from which 7 previously reported candidate proteins that involved in abiotic stresses were selected and cloned for rotated verification in yeast. As a result，one upstream signal regulator of ABA pathway，VaPYL9 was preliminary confirmed to interact VaCIPK18. Furthermore，Y2H rotation verification and bimolecular fluorescence complementation（BiFC）were used to verify the interaction between them. Our result demonstrated that VaPYL9 specifically interacts with VaCIPK18 and VaCIPK18△NAF protein in yeast，and VaPYL9 could also interact with VaCIPK18△S_TKc and VaCIPK18△NAF protein in plant cells. Interestingly，there is no interaction between VaPYL9 and VaCIPK18 in Arabidopsis protoplast.
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• Expression Analysis of MiPGP1 and MiPGP2 and Their Effects on Adventitious Root Formation in Arabidopsis
• LI Yunhe, LIU Min, LIU Jiandong, and WU Qingsong
• Acta Horticulturae Sinica. 2020, 47(12): 2317-2330. DOI:10.16420/j.issn.0513-353x.2020-0422
• Abstract ( 228 ) HTML ( 192 ) PDF (2119KB) ( 192 ) 　　　
• Adventitious roots form only at the proximal cut surface（PCS）but not at the distal cut surface（DCS）of mango cotyledon segments，it was suggested that polar auxin transport play important role in this process but the role of multi-drug-resistant/P-glycoprotein（MDR/PGP）in this rooting was unclear. In this study，two mango MDR/PGP genes，designated as MiPGP1 and MiPGP2，were isolated from the cotyledon of mango（Mangifera indica L.‘Yuexi 1’）using RT-PCR and RACE. The full-length cDNA of MiPGP1 and MiPGP2 were 4 650 and 4 107 bp，with the open reading frames of 4 092和3 851 bp，which encode two putative proteins of 1 363 and和1 289 amino acids，respectively. The genomic DNA sequences of MiPGP1 and MiPGP2 were 6 430 and 5 747 bp，with the lengths of 6 082 and 5 633 bp，respectively，from the start codon to the terminator codon. Phylogenetic tree analysis indicated that MiPGP1 and MiPGP2 belong to plant PGP1 group. Quantitative real-time PCR showed that MiPGP1 and MiPGP2 expressed in PCS（where adventitious root formed）or DCS（no adventitious root formed）throughout the adventitious root formation period. Meanwhile，the relative expression of MiPGP1 and MiPGP2 was low at 0 d，but which was significantly up-regulated from 6 h until adventitious root appeared at 10 d，and the relative expression was always higher in DCS than that of PCS. However，the relative expression of MiPGP2 was significantly up-regulated by the pre-treatment with indole-3-butyric acid（IBA）but not MiPGP1. Pre-treatment with 2,3,5-triiodobenzoic acid（TIBA）inhibited the relative expression of MiPGP1 and MiPGP2 at some time points. In additon，the results of MiPGP1 and MiPGP2 overexpression in Arabidopsis plants showed that there was a significant increasment in adventitious root quantity in MiPGP2 transgenic line in hypocotyl. In conclusion，MiPGP2 might play an important role during the adventitious root formation of mango cotyledon segments.
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• FaRGA1 Gene Silencing Changes the Characteristics of Flowering and Runner Producing in Strawberry
• LUO He, LI Weijia, LI He, and ZHANG Zhihong
• Acta Horticulturae Sinica. 2020, 47(12): 2331-2339. DOI:10.16420/j.issn.0513-353x.2020-0584
• Abstract ( 450 ) HTML ( 278 ) PDF (4430KB) ( 278 ) 　　　
• RGA is one of the DELLA protein that is the core component of inhibition of gibberellin-mediated response in the gibberellin signaling pathway，and plays an important role in plant growth and development. In this study，the physicochemical characteristics and secondary structure of FaRGA1 in cultivated strawberry（Fragaria × ananassa）were predicted and the conserved domain of FaRGA1 protein was analyzed. Phylogenetic tree analysis shows that FaRGA1 has high homology with FveRGA1 in woodland strawberry（F. vesca），and the two are orthologous genes. The RNAi vector of strawberry RGA1 gene was constructed，and 3 transgenic lines with FaRGA1 silencing were obtained from strawberry cultivar‘Jingyu’by Agrobacterium-mediated genetic transformation. Compared with non-transgenic control plants，all three FaRGA1 silencing lines showed the characteristics of without flowers and continuously producing runners during two years，and the shortened crowns were significantly elongated and lignified. This study found that FaRGA1 gene has an inhibitory effect on tunner formation and promotes flowering in cultivated strawberries，which lays the foundation for molecular breeding of strawberry based on FaRGA1 gene in the future.
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• Characterization and Genetic Analysis of a Yellowing Mutant of Eggplant Leaf Color
• LIU Fuzhong, ZHANG Ying, YANG Jinkun, CHEN Yuhui, SHU Jinshuai, LI shupei, and CHEN Lulu
• Acta Horticulturae Sinica. 2020, 47(12): 2340-2348. DOI:10.16420/j.issn.0513-353x.2020-0829
• Abstract ( 475 ) HTML ( 311 ) PDF (1249KB) ( 311 ) 　　　
• A leaf color yellowing mutant‘chl861-2’and its normal leaf near-isogenic line ‘0643-1’were used to study their leaf color change trend，chlorophyll content and several agronomic traits. The mutant had a chlorophyll-deficient characteristic. Its leaf color yellowing phenomenon was exhibited since the cotyledon stage and throughout the growth period. Its cotyledon was light yellow，true leaf color was yellow. The mutant plants grew slowly，dwarfed，and prolonged their growth period. The fruits of mutant plants were significantly smaller than those of the wild type，and the fruit weight was about 71.58% of that of the wild type. The contents of total chlorophyll，chlorophyll a and chlorophyll b in the mutant were significantly lower than those in the wild type at the seedling stage，the first flowering stage，and the fourth fruiting stage. The net photosynthetic rate（Pn）of the mutant was significantly lower than that of the wild type at the seedling stage and the flowering stage，which stomata conductance（Gs）and transpiration rate（Tr）were lower than those of the wild type，and its intercellular CO2 concentration （Ci）is higher than the wild-type. The inheritance of the leaf color yellowing trait of‘chl861-2’was analyzed by using the segregating populations derived from the crosses between‘chl861-2’and three normal leaf color inbred lines. The results showed that the leaf color yellowing trait was controlled by a recessive nuclear single gene，which was named Smchl-1.
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• HS–SPME–GC–MS Analysis of Volatile Components in Tender Shoots from Six Plants of Calycanthaceae
• SHEN Zhiguo, SUN Meng, YUAN Deyi, CHENG Jianming, DING Xin, and SHANG Zhonghai,
• Acta Horticulturae Sinica. 2020, 47(12): 2349-2361. DOI:10.16420/j.issn.0513-353x.2020-0507
• Abstract ( 223 ) HTML ( 203 ) PDF (1754KB) ( 203 ) 　　　
• The headspace solid phase microextraction（HS–SPME）combained with the GC–MS test were used to detect the volatile components of six Calycanthaceae plants. Moreover，in order to investigate the volatile components difference of the tender shoots between several species（strains）of Calycanthaceae，the common component analysis，principal component analysis and cluster analysis of the volatile components were adopted to compare the Chimonanthus nitens（a widely used medicinal plant）with other tested species（strains）. The results showed that a total of 141 volatile components were detected in the tender shoots, with 47 in Ch. praecox‘Hongyun’，48 in Ch. praecox‘Jinyu’，44 in Ch. praecox‘Yanlingsuxin’，44 in Ch. nitens，48 in Calycanthus chinensis and 46 in Ca. floridus. Alkenes and alcohols were the main volatile components in the six plants. Moreover，when compared with Ch. nitens，the common volatile components of Ch. praecox‘Hongyun’，‘Jinyu’，‘Yanlingsuxin’，Ca. chinensis，and Ca. floridus were 15，18，10，14 and 20，respectively. PCA analysis and cluster analysis showed that Ca. chinensis falled into a group，while Ch. nitens and Ch. floridus belong to one group，and Ch. praecox‘Hongyun’，‘Jinyu’and‘Yanlingsuxin’are the other one. Furthermore，we found eudesmol and β-terpinene were the primary differential volatile compounds when executing the comparative analysis with Ch. nitens. Overall，abundant volatile components were found in tender shoots of the six Calycanthaceae plants，and our study have great potential for medical development though there are certain discrepancy among different species（strains）
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• Relationship Between the Composition and Content of Anthocyanin in Petals of Ranunculus asiaticus and Anemone cathayensis and Their Flower Color
• MENG Han, FU Dengxiang, WU Yanmei, and JIN Xuehua
• Acta Horticulturae Sinica. 2020, 47(12): 2362-2372. DOI:10.16420/j.issn.0513-353x.2020-0166
• Abstract ( 436 ) HTML ( 264 ) PDF (1131KB) ( 264 ) 　　　
• This study uses 10 different color Ranunculus asiaticus and 4 different color Anemone cathayensis as the materials. It measures the flower color phenotype of petals with visual colorimetry method，RHSCC colorimetric card colorimetry and colorimeter（CR-400），and determines the composition and structure of anthocyanins in petals with UV-vis spectrophotometer（TU-1901）and high performance liquid chromatography-electrospray ionization-mass spectrometry（HPLC–ESI–MS）. And then it analyzes the relationship between flower color and anthocyanin composition with multiple linear regression. The results shows that five anthocyanins are detected in the petals of eight anthocyanin accumulating varieties of Ranunculus asiaticus，namely，pelargonidin，cyanidin，delphinidin，peonidin and malvidin. These anthocyanins are further modified to form 15 anthocyanins. The red and pink varieties mainly contain peonidin and cyanidin，while in yellow variety，pelargonidin or carotenoid is the main anthocyanin component. According to the analysis，the contents of cyanidin glycoside and pelargonidin glycoside are positively correlated with L* value，and the accumulation of pelargonidin glycoside has a greater contribution to petal brightness. Six anthocyanin are detected in the petals of four varieties of Anemone cathayensis，namely，petunidin，pelargonidin，cyanidin，delphinidin，peonidin and malvidin. They form 20 anthocyanins by different glycosylation and acylation modification，showing rich anthocyanin diversity.
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• RNAi Silencing CsHB1 Reduces the Accumulation of Caffeine in Tea Callus
• KANG Xin, LIU Ping, MA Wenhui, ZHANG Yuanyuan, LIN Xiaorong, LI Bin, and CHEN Zhongzhen,
• Acta Horticulturae Sinica. 2020, 47(12): 2373-2384. DOI:10.16420/j.issn.0513-353x.2020-0242
• Abstract ( 271 ) HTML ( 215 ) PDF (2566KB) ( 215 ) 　　　
• In this study，the HD-Zip transcription factor CsHB1 gene was cloned from‘Yinghong 9’（Camellia sinensis）tea plant，and the recombinant CsHB1 protein was expressed in Escherichia coli. The subcellular localization of CsHB1 was carried out using tobacco as an expression host. The RNAi transgenic vector of CsHB1 was constructed and transformed into‘Yinghong 9’tea callus mediated by Agrobacterium，followed by the analysis of gene expression and caffeine content on the silence CsHB1 callus. Our results showed that the cloned CsHB1 gene shared the same amino acid sequence exception 6 nucleotides difference with NCBI registered CsHB1 gene（accession No. MF033534）isolated from‘Longjing 43’tea plant. A 49 kD insoluble recombinant protein corresponding to the CsHB1 gene was obtained by prokaryotic induction. The expression protein infused with GFP as GFP-CsHB1 located in the cell nucleus and cytoplasm of tobacco. In the silence expression of CsHB1 callus，the expression of the CsHB1 gene decreased significantly，and the expression of yhNMT1，an N-methyltransferase，which catalyzes the synthesis of caffeine in tea，was also found decreased by real-time fluorescence quantitative PCR analysis. HPLC analysis showed that the content of caffeine in the transgenic callus decreased markedly. Based on these results，we suggest that the silencing of CsHB1 suppresses the expression of yhNMT1，and then reduces the accumulation of caffeine in the callus of‘Yinghong 9’.
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• Establishment of Genetic Transformation System of Lycium barbarum Endophytic Fungus Fusarium nematophilum NQ8GⅡ4 Using PEG-mediated Method and Transformants Evaluation
• YAN Siyuan, DU Juan, YANG Fulong, REN Miaomiao, LI Jiahong, and GU Peiwen
• Acta Horticulturae Sinica. 2020, 47(12): 2385-2396. DOI:10.16420/j.issn.0513-353x.2020-0131
• Abstract ( 227 ) HTML ( 283 ) PDF (6251KB) ( 283 ) 　　　
• In this study，the expression vector of green fluorescent protein was transferred into the strain NQ8GⅡ4 by PEG-mediated protoplast transformation. Transformants of no difference with the wild strain were obtained by biological assay and PCR analysis. The cell walls of young hypha of NQ8GⅡ4 strain was lyzed with enzymes to generate protoplasts，which were used to insert the exogenous DNA randomly under PEG8000 mediated transformation. Fifty-seven transformants of NQ8GⅡ4 that showed hygromycin B resistance and fluorescence signal were obtained. The transformation frequency of NQ8GⅡ4 was 2 850 transformants per mg DNA. The transformants were compared with wild type strain for mitotic stability，phenotype，fluorescence，antagonistic activity and pathogenicity and a transformant with similar properties to wild-type strain was screened out. The results have established foundation for futher study on the endogenesis and biocontrol mechanism of strain NQ8GⅡ4.
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Research Notes

• The Disease Development of Apple Scar Skin Viroid in the Field and Its Transmission Mode in Tissue Culture
• XI Nana, ZHAO Ke, YANG Jinfeng, MENG Xianglong, HU Tongle, WANG Shutong, WANG Yanan, and CAO Keqiang
• Acta Horticulturae Sinica. 2020, 47(12): 2397-2404. DOI:10.16420/j.issn.0513-353x.2020-0068
• Abstract ( 240 ) HTML ( 220 ) PDF (1502KB) ( 220 ) 　　　
• Apple scar skin disease caused by apple scar skin viroid（ASSVd）is common in apple producing areas northern in China and it has led to serious economic losses in recent years. At present，there are no effective treatment measures. In order to clarify the disease development and transmission mode in the field，three orchards in Nanshennan Village，Shunping，Baoding were continuously investigated and tested from 2012 to 2015. In orchard A，B，C，the change of symptom rate from 3.00%，19.35% and 10.00% to 10.00%，34.41% and 22.00%，and the change of virus-taking rate from 11.00%，20.43% and 14.00% to 23.00%，37.63% and 30.00%. The rate of symptom-showing and virus-taking increased year by year，and the virus-taking rate was greater than the symptom-showing rate. The incidence of fruit trees in the field distribution has an obvious incidence center. Some fruit tree taking virus did not show symptom. By high-throughput sequencing method，it was found that the number of ASSVd vsiRNA reads in symptomatic fruit trees was 4.08 times that of asymptomatic fruit trees with virus，and the accumulation of ASSVd in symptomatic fruit trees was significantly higher than that in asymptomatic fruit trees with virus by quantitative real-time PCR detection. ASSVd-carrying tissue culture seedlings and non-ASSVd- carrying tissue culture seedlings were used as test materials to simulate different transmission modes in the field，and RT-PCR was used to confirm that ASSVd could be transmitted through wound contamination，tissue culture shear pollution，virus-taking medium and root contact，among which the root contact transmission rate was the highest.
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• Cloning and Expression Analysis of CsNCED3-2 in Responding to Citrus Canker Disease
• XIE Yu, ZHANG Qingwen, QI Jingjing, ZOU Xiuping, HE Yongrui, XU Lanzhen, LEI Tiangang, PENG Aihong, LI Qiang, YAO Lixiao, CHEN Shanchun, and LONG Qin
• Acta Horticulturae Sinica. 2020, 47(12): 2405-2414. DOI:10.16420/j.issn.0513-353x.2020-0170
• Abstract ( 293 ) HTML ( 263 ) PDF (3896KB) ( 263 ) 　　　
• Citrus canker is an important disease of citrus in the world and causes severe economic losses to the citrus industry. In this study，the 9-cis-epoxycarotenoid dioxygenase gene（NCED）family of Citrus sinensis were annotated，and the citrus canker related CsNCED3-2 was cloned and its expression pattern was analyzed. A total of 14 NCED family members were annotated from the public genomic databases of C. sinensis. These 14 NCED family members can be divided into 4 subclasses based on the phylogenetic tree and functional domain. The full-length of CsNCED3-2 is 2 377 bp with a 1 830 bp open reading frame which codes a protein containing 609 amino acids. The CsNCED3-2 gene belongs to the members of the RPE65 superfamily and the carotenoid dioxygenase family，and it is a non-secretory protein. After inoculated with Xanthomonas citri subsp. citri（Xcc），the expression of CsNCED3-2 in Jindan（Fortunella crassifolia Swingle）was always higher than that in Wanjincheng orange（Citrus sinensis Osbeck）at different time points，and the variation trends of expression in the two varieties were opposite. The expression of CsNCED3-2 in the peel of Wanjincheng Orange and Jindan were higher than that in the stem，leaf and seed. The CsNCED3-2 promoters of Wanjincheng Orange and Jindan contain multiple cis-acting elements involved in plant adversity and hormone responses，such as ABRE（abscisic acid response），TCA-element（salicylic acid response），MYC/MYB（drought response）and so on，but there are differences in their number and type.
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• Identification of Pepper Vein Yellows Virus and Evolution Analysis of P0，CP and MP Genes
• WANG Lishuang, LI Chun, YANG Xuehui, ZHANG Songbai, and LIU Yong,
• Acta Horticulturae Sinica. 2020, 47(12): 2415-2426. DOI:10.16420/j.issn.0513-353x.2020-0167
• Abstract ( 366 ) HTML ( 220 ) PDF (1467KB) ( 220 ) 　　　
• In order to find out the occurrence and distribution of pepper vein yellows virus（PeVYV）and its molecular variation on pepper in Guizhou. 548 suspected samples from 25 sampling location in Guizhou Province were analyzed using RT-PCR. The results showed that 129 samples were positive and the detection rate was 23.54%. The sequence alignment of the PeVYV isolates from 14 different locations with the reported PeVYV isolates from China and other countries was performed，the nucleotide and amino acid homology of P0 gene was 90.9%–100.0%，amino acid similarity was 97.7%–100.0%，the nucleotide and amino acid homology of CP gene was 93.7%–100.0%，amino acid similarity was 97.9%–100.0%，the nucleotide and amino acid homology of MP gene was 94.7%–100.0%，amino acid similarity was 100.0%. In the phylogenetic tree，the results indicated that P0 of PeVYV from 13 locations were clustered and closely related to the isolates（KP326573），isolates from Bozhou，Zunyi，were closely related to the isolates from Australia. All PeVYV isolates CP gene in Guizhou were clustered with isolates from China，Australia，the United States and Spain. But the PeVYV isolates from different locations were clustered into two different branches. All MP genes from different locations were clustered together，and there was no significant difference in comparing with isolates from the other regions. The research results show that PeVYV isolates from different regions had molecular variation and genetic differentiation.
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• Induction and Plant Regeneration of Virus-free Microtuber in Red Bud Taro
• LIU Xingyue, ZHU Qianglong, LI Huiying, SUN Jingyu, LI Bicong, Zhang Hongyu, HUANG Yingjin, FANG Jiajun, and ZHOU Qinghong
• Acta Horticulturae Sinica. 2020, 47(12): 2427-2438. DOI:10.16420/j.issn.0513-353x.2020-0470
• Abstract ( 294 ) HTML ( 276 ) PDF (2718KB) ( 276 ) 　　　
• In order to solve the low survival rate and high differentiation rate of virus-free seedling of red bud taro seedlings in the process of transplanting，in this study，with taking the most famous and high-quality local cultivars of Yanshan red bud taro（Colocasia esculenta）in Jiangxi Province as experimental material，the detoxification culture of taro stem tip，molecular detection of test-tube plantlets and microtuber induction were carried out. The results showed that seedlings in vitro could be induced from 0.2–1.0 mm taro buds inoculated in MS + 2.0 mg • L-1 6-BA + 0.5 mg • L-1 NAA + 3% sucrose medium，and RT-PCR molecular detection indicated that the seedlings induced from stem tip under 0.3 mm diameter in culture could not carry virus（DsMV）. The optimal medium for the induction of secondary proliferation of virus-free seedlings was MS + 3.0 mg • L-1 6-BA + 0.5 mg • L-1 NAA + 3% sucrose，the optimal medium for the induction of virus-free cormel was MS + 1.0 mg • L-1 6-BA + 0.5 mg • L-1 NAA + 8% sucrose，the optimal culture virus-free seedlings and taro corm conditions were 25 ℃，54 μmol • m-2 • s-1，14 h • d-1，the optimal substrates for transplanting and acclimation of was peat︰vermiculite（2︰1），and the survival rate was 97.71%. Compared with the contrast，the yield of virus-free taro increased 43.14% and there were significant differences in nutritional characters between them. The formation of virus-free cormel and the establishment of a plant regeneration system provide a new way for the rapid propagation of healthy seedlings of red bud taro.
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• Complete Chloroplast Genome Sequence of Clivia miniata and Its Characteristics
• ZHENG Yi, ZHANG Hui, WANG Qinmei, GAO Yue, ZHANG Zhihong, and SUN Yuxin
• Acta Horticulturae Sinica. 2020, 47(12): 2439-2450. DOI:10.16420/j.issn.0513-353x.2020-0466
• Abstract ( 346 ) HTML ( 289 ) PDF (1560KB) ( 289 ) 　　　
• The total DNA of Clivia miniata leaves was sequenced by Illumina MiSeq sequencing platform，and the full-length sequence（158 114 bp）of chloroplast genome（cpDNA）was obtained by assembly. A total of 135 genes were annotated in the cpDNA. The 135 genes included 87 protein coding genes，40 tRNA genes and 8 rRNA genes. Both simple sequence repeat（SSR）and codon preference of the cpDNA were analyzed using bioinformatics method. The results showed that（1）the total number of SSR loci in the cpDNA of C. miniata was 61，of which 38，9，2，8，3 and 1 were mononucleotide，dinucleotide，tirnucleotide，tetranucleotide，pentanucleotide and hexanucleotide repeats，respectively，and most of the SSRs were located in the intergenic regions；（2）the codons in cpDNA of C. miniata tended to end with A or U(T)，the frequency of serine was the highest and that of cysteine was the lowest. In addition，the phylogenetic analysis was carried out based on the full sequence of 24 cpDNAs and the sequence of 23 ycf2 genes of chloroplasts. The results showed that both C. miniata and Amaryllidaceae plants were clustered into one branch，which not only indicated the closest genetic relationship of them but also supported that C. miniata belongs to Amaryllidaceae. The results of phylogenetic analysis based on chloroplast ycf2 are mostly the same as that based on full sequence of cpDNA，which indicated that the ycf2 genes could replace full-length cpDNAs for plant phylogenetic analysis.
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New Technologies and New Methods

• Efficient Identification of Tetraploid Plants from Seedling Populations of Apomictic Citrus Genotypes Based on Morphological Characteristics
• ZHOU Rui, XIE Kaidong, WANG Wei, PENG Jun, XIE Shanpeng, HU Yibo, WU Xiaomeng, and GUO Wenwu,
• Acta Horticulturae Sinica. 2020, 47(12): 2451-2458. DOI:10.16420/j.issn.0513-353x.2020-0769
• Abstract ( 495 ) HTML ( 257 ) PDF (1109KB) ( 257 ) 　　　
• Based on the phenomenon of spontaneous doubling of nucellar cells in polyembryonic citrus genotypes，we established a simple and efficient approach for citrus tetraploid exploitation by sowing the seeds（after germination being accelerated in a 28 ℃ incubator for about one week）in pots in a controlled growth chamber，followed by screening tetraploids from germinated seedlings based on root，leaf morphology，oil gland density and flow cytometry analysis. By this method，we screened 5，6，5，17，45 and 7 putative tetraploids respectively from the 432，506，1 356，1 870，2 749 and 685 seedlings of Succari sweet orange，Hongjiang sweet orange，Gonggan mandarin，Nian mandarin，Xinhui mandarin and Huapi kumquat. By analyzing their ploidy level using flow cytometry，a total of 72 tetraploids was obtained，including 4 from Succari sweet orange，5 from Gonggan mandarin，5 from Hongjiang sweet orange，14 from Nian mandarin，39 from Xinhui mandarin and 5 from Huapi kumquat，with the tetraploid pre-screening accuracy rate of 80.0%，83.3%，100.0%，82.4%，86.7% and 71.4%，respectively. The merits of our new method are higher accuracy rate of tetraploid pre-screening，and time saving only needing less than 40 d from seed sowing to tetraploid confirmation. Simple sequence repeat（SSR）analysis showed that the bands of all tetraploids obtained herein are identical with that of their corresponding diploid genotypes，indicating that all these plants are the doubled diploids which have great potential for seedless triploid breeding.
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New Cultivars

• A New Anthurium andraeanum Cultivar‘Fendai’
• TIAN Danqing, GE Yaying, PAN Xiaoyun, PAN Gangmin, ZHU Qiang, ZHOU Yuan, and WAN Xiao
• Acta Horticulturae Sinica. 2020, 47(12): 2459-2460. DOI:10.16420/j.issn.0513-353x.2020-0125
• Abstract ( 314 ) HTML ( 118 ) PDF (1138KB) ( 118 ) 　　　
• Anthurium andraeanum‘Fendai’is a new potted cultivar from the cross between‘Sempre’（female parent）and‘Big Beauty’（male parent）. It is a medium plant with average plant height of 32.5 cm. The leaves are ovate with average length of 17.3 cm and width of 10.5 cm. The spathes are higher than leaves，showing broad ovate，dark pink，and extremely strong depression，being 9.9 cm in length and 8.8 cm in width. Spadixes are up-right and red with average length of 3.8 cm. This cultivar grows fast，performs well in winter，has many flowers and is bright in color.
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• New Primulina Cultivars‘Lucky’and‘Yuyan’
• YAN Haixia, SONG Qian, TAO Dayan, ZHOU Jinye, GUAN Shikai, LI Xiuling, and HE Jingzhou
• Acta Horticulturae Sinica. 2020, 47(12): 2461-2462. DOI:10.16420/j.issn.0513-353x.2019-0932
• Abstract ( 277 ) HTML ( 136 ) PDF (1101KB) ( 136 ) 　　　
• Primulina‘Lucky’is selected from the hybrid（P. glandaceistriata × P. sclerophylla），‘Yuyan’is selected from the hybrid （P. liujiangensis × P. glandaceistriata）. These hybrids are perennial herbaceous rosette plants，and the flowering period is from April to May. It is easy to cultivate and propagate. Compared with the parents，the lower lip of the corolla of‘Lucky’is very long；the leaves of‘Yuyan’have white reticulated veins，and the flowers are large，which has the characteristics of viewing flowers and leaves.
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• A New Landscaping Xanthoceras sorbifolium Cultivar‘Senmiao Jinfenguan’
• WANG Yali, JIA Qiaoxia, and LI Yi,
• Acta Horticulturae Sinica. 2020, 47(12): 2463-2464. DOI:10.16420/j.issn.0513-353x.2019-0696
• Abstract ( 786 ) HTML ( 121 ) PDF (1378KB) ( 121 ) 　　　
• Xanthoceras sorbifolium‘Senmiao Jinfenguan’is a new cultivar selected from seedling. It is high of tree vigor. The petal color constantly changed with the prolonging of blossom. The petal from middle to edge is lemon in the bud period gradually change to light yellow in the early blossom then gradually change to light pink in the middle and late blossom. The petal base is yellow gradually change to pink then change to red. The margin of petal fringed. The fruit is peach shaped. The mature seed is brown and the weight of 1 000 seeds is 920 g. The seed number in each capsule is 13. The suitable cultivation area is Ningxia，Inner Mongolia，Shaanxi，Gansu and other main planting area of Xanthoceras sorbifolium.
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