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Lili Dong, Qi Wang, Feng Xiong, Na Liu and ShuiMing Zhang

More axillary buds 1 (MAX1), initially identified in arabidopsis (Arabidopsis thaliana), is a key regulatory gene in strigolactone synthesis. CmMAX1, an ortholog of MAX1 was cloned from chrysanthemum (Chrysanthemum morifolium cv. Jinba). It had an open reading frame of 1611 bp and encoded 536 amino acid of P450 protein, with a conserved heme-binding motif of PFG × GPR × C × G, as well as PERF and KExxR motifs. The predicted amino acid sequence of CmMAX1 was most closely related to the MAX1 ortholog identified in lotus (Nelumbo nucifera), NnMAX1, with 55.33% amino acid sequence similarity. Expression analysis revealed there was no significant difference of CmMAX1 expression among various tissues. Phosphorus (P) deficiency significantly improved the expression levels of CmMAX1. Strigolactone, auxin, and cytokinin negatively regulated the expression of CmMAX1. Overexpression of CmMAX1 reduced the branch numbers of arabidopsis max1-1. These results suggest that CmMAX1 may be a candidate gene for reducing the shoot branching of chrysanthemum.

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Weisheng Liu, Dongcheng Liu, Aimin Zhang, Chenjing Feng, Jianmin Yang, Jaeho Yoon and Shaohua Li

Inter-simple sequence repeat (ISSR) markers were used to evaluate genetic similarity and interrelationship among 104 plum (Prunus L. spp.) and related accessions from the Chinese National Germplasm Repository for Plums and Apricots and the Tianshan Germplasm Repository for Wild Fruit Resources, including six plum species (Prunus salicina Lindl., Prunus simonii Carr., Prunus ussuriensis Kov. et Kost., Prunus domestica L., Prunus cerasifera Ehrh., and Prunus spinosa L.), two related species [apricot (Prunus armeniaca L.) and nanking cherry (Prunus tomentosa Thunb.)], eight putative hybrids between plum and apricot (plumcot), and six accessions of wild European plum (P. domestica). Out of the 42 ISSR primers, 12 were selected, which generated 103 markers in total, 99 of which were polymorphic. Possible accession-specific ISSR bands or patterns were also found. Some possible synonyms or homonyms were clarified or discussed, and closely related accessions such as bud mutants were discriminated. Based on the unweighted pair group method with arithmetic mean (UPGMA) analysis and principal coordinate analysis (PCoA) using the Jaccard coefficient, two different dendrograms were constructed—one including accessions grouped by species and one with all 104 accessions—and a two-dimensional plot was obtained. Three groups were formed in both dendrograms and PCoA plot: Group I including apricot (‘Yinxiangbai’) and plumcot types; Group II containing Asia-originated diploid species [e.g., P. cerasifera, P. ussuriensis, P. tomentosa, and Chinese plum-types (i.e., P. salicina and its hybrids)]; and Group III involving European-origin polyploid species (e.g., P. spinosa and P. domestica) and recently found wild European plum accessions in China. The dendrogram with accessions grouped by species implied that 1) plumcot types had closer relatedness with apricot than with plum; 2) P. simonii should be a variant of P. salicina while P. ussuriensis an independent species; 3) P. domestica was more closely related to P. spinosa than to P. cerasifera. Two accessions of European plum (‘89-7-3’ and ‘Wanhei’) were clustered into outgroups in the dendrogram with all 104 accessions, which could been grouped within Group III in the PCoA plot. The distribution of both European plum and Chinese plum-types across respective groups did not reflect the geographic origins. The present study also further confirmed that the wild plants found in Xinjiang of China were P. domestica.

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Xiaoxu Yang, Chang Liu, Zhishan Yan, Youjun Fan, Guojun Feng and Dajun Liu

Flowering time influences pod yield and quality of common bean (Phaseolus vulgaris); however, our knowledge of flowering time genes and flowering mechanisms in common bean remain limited. We performed RNA-sequencing (RNA-seq) analyses [long-day (LD) condition and short-day (SD) condition] to identify the flowering time genes and analyzed differentially expressed genes to examine their expression levels in relation to flowering time in ‘Hong Jin Gou’ common bean, a cultivar highly sensitive to photoperiod. The circadian patterns of related genes were identified using quantitative real-time polymerase chain reaction (qRT-PCR). Flowering time in ‘Hong Jin Gou’ was influenced by day length: SD conditions promoted flowering. A total of eight flowering time–related genes were identified, which were classified into photoperiod pathways. Homologs of pseudo-response regulator 5, pseudo-response regulator 7, and gigantea were more highly expressed under SD conditions than under LD conditions. Homologs of late elongated hypocotyl and timing of cab expression 1 were differentially expressed under light and dark conditions. Early flowering 3 is a key regulator of the pathway, which coordinates light and circadian clock inputs in leaves to trigger the expression of downstream genes. The present study provides critical information that could facilitate further investigations on the genetic mechanism of flowering time in common bean.

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Jian-rong Feng, Wan-peng Xi, Wen-hui Li, Hai-nan Liu, Xiao-fang Liu and Xiao-yan Lu

The characterization of aroma of the 14 main apricot (Prunus armeniaca L.) cultivars in Xinjiang was evaluated using high-performance solid-phase microextraction (HP-SPME) with gas chromatography-mass spectroscopy (GC-MS). A total of 208 volatiles that include 80 esters, 25 aldehydes, 15 terpenes, 21 ketones, 39 alcohols, 27 olefins, and 1 acid were identified from these cultivars. The compounds propyl acetate, 3-methyl-1-butanol acetate, (Z)-3-hexen-1-ol acetate, d-limonene, β-linalool, hexanal, hexyl acetate, butyl acetate, β-myrcene, ethyl butanoate, and β-cis-ocimene were the major compounds responsible for aroma in these cultivars. GC-MS results showed that Kuchexiaobaixing, Guoxiyuluke, and seven other cultivars were characterized by a high level of esters and were considered to be fruity apricot aroma. ‘Luotuohuang’ and ‘Heiyexing’ accumulate high levels of terpenes and exhibited an outstanding floral aroma. Higher levels of alcohols and aldehydes were observed in ‘Danxing’, ‘Sumaiti’, and ‘Kumaiti’. The latter are considered green aroma cultivars. These three types of cultivars with different aroma characteristics can be significantly differentiated by using the principal component analysis (PCA) method. The contributions of volatiles to the apricot aroma were assessed by using the partial least squares regression (PLSR) model. Esters, terpenes, and C6 components were shown to be responsible for the fruity, floral, and green character of fresh apricots, respectively.

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Hai-nan Liu, Jian-rong Feng, Xiao-fang Liu, Wen-hui Li, Wen-juan Lv and Ming Luo

Three kinds of expression vectors of a pollen-S determinant were constructed to provide a reference for molecular breeding of self-compatible (SC) Prunus species. An S-haplotype-specific F-box (SFB) protein gene from the ‘Xiaobaixing’ apricot (Prunus armeniaca) was cloned by reverse transcription polymerase chain reaction (RT-PCR) and 3′-rapid-amplification of cDNA ends (3′-RACE). A 1136-bp sequence complementary to the 3′-end of the cDNA (GenBank accession number KP938528.2) with a 912-bp complete open reading frame (ORF) was obtained. The deduced amino acid sequence contained an F-box domain, two variable regions, and two hypervariable regions with structural characteristics similar to SFB in other Rosaceae plants. Sense, antisense, and RNA interference (RNAi) vectors for SFB were constructed by enzyme restriction. The target fragment was restricted using the corresponding restriction enzyme and then directionally inserted between the 35S cauliflower mosaic virus promoter and the nopaline synthase terminator (NOS-ter) of the expression vector pCAMBIA-35S-MCS-NOS-NPTII. The intron-containing hairpin RNA (ihpRNA) was obtained by fusion PCR. The constructed vectors were transferred into Agrobacterium tumefaciens strain LBA4404 by freezing/thawing. The RNAi vector of SFB was also transformed in tobacco (Nicotiana tabacum). The successful construction of these three expression vectors provides a basis for transforming ‘Xiaobaixing’ apricot and the breeding of SC Prunus cultivars.

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Feng Gao, Arvind H. Hirani, Jun Liu, Zheng Liu, Guohua Fu, Chunren Wu, Peter B.E. McVetty and Genyi Li

There are various clubroot pathogen (Plasmodiophora brassicae) resistance genes within Brassica species with european turnip (B. rapa ssp. rapifera) being identified as potentially the best source of resistance for the development of clubroot-resistant cultivars in chinese cabbage (B. rapa ssp. pekinensis). To use clubroot resistance genes effectively, it is necessary to map these genes so that molecular markers inside or closely linked to these resistance genes can be developed. Using molecular marker-assisted selection, the clubroot resistance genes can be effectively transferred from cultivar to cultivar and from species to species. In this report, one clubroot resistance locus was mapped on linkage group A3 using five segregating populations developed from five chinese cabbage cultivars, suggesting that all the five cultivars shared the same clubroot resistance locus. Furthermore, one of these five chinese cabbage cultivars was used to develop a large segregating population to fine-map this clubroot resistance locus to a 187-kilobp chromosomal region. Molecular markers that are closely linked to the mapped clubroot resistance locus have been developed that can be used for marker-assisted selection in chinese cabbage and canola/rapeseed (B. rapa and B. napus) breeding programs.

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Chao Gao, Deyi Yuan, Ya Yang, Bifang Wang, Dongming Liu and Feng Zou

Camellia oleifera is an important plant species that produces edible oils. Understanding the double fertilization of this plant is critical for studies concerning crossbreeding, self-incompatibility, and the biological mechanisms underlying hybridization. We aimed to characterize pollen tube growth and double fertilization in C. oleifera. The female and male parent cultivars (Huashuo and Xianglin XLC15, respectively) were used for artificial pollination. Growth of the pollen tube in the style, ovary, and ovule from pollination to fertilization and the cytological characteristics of female and male gamete fusion during double fertilization were observed using fluorescence and scanning electron microscopy (SEM). Numerous pollen grains germinated 2 to 4 hours after pollination. The pollen tubes entered the interspaces between the papillar cells, grew along the stylar canal, and aggregated at the one-third site of the style. They grew in the gradually narrowing stylar canal, entering the locule. The tubes turned 90° and entered the embryo sac through the micropyle; subsequently, they entered a degenerated synergid, where the spermatids were released. One sperm nucleus fused with the polar nucleus, forming the primary endosperm nucleus, whereas the other sperm fused with the egg, forming the zygote. The polar nucleus was fertilized earlier than the egg. Double fertilization of C. oleifera is characterized as pre-mitotic gametogony. The current results lay a theoretical foundation for studies concerning the crossbreeding and embryology of C. oleifera and provide fundamental data concerning the reproductive biology of the genus Camellia.

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Kai Zhao, Feng Zhang, Yi Yang, Yue Ma, Yuexue Liu, He Li, Hongyan Dai and Zhihong Zhang

GA20-oxidase (GA20-ox) is a key enzyme involved in the biosynthesis of gibberellic acid (GA). To investigate its role in plant growth and development, we suppressed MdGA20-ox gene expression in apple (Malus domestica cv. Hanfu) plants by RNA interference (RNAi). After 20 weeks of growth in the greenhouse, significant phenotype differences were observed between transgenic lines and the nontransgenic control. Suppression of MdGA20-ox gene expression resulted in lower plant height, shorter internode length, and higher number of nodes compared with the nontransgenic control. The expression of MdGA20-ox in transgenic plants was significantly suppressed, and the active GA content in transgenic lines was lower than that in the nontransgenic control. These results demonstrated that the MdGA20-ox gene plays an important role in vegetative growth, and therefore it is possible to develop dwarfed or compact scion apple cultivars by MdGA20-ox gene silencing.

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Yuting Meng, Boling Liu, Ping Zhang, Ping Cui, Yuguang Song, Nianwei Qiu, Guoliang Han and Feng Zhou

2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) is one of the most toxic polybrominated diphenyl ethers (PBDEs). The toxic effects of BDE-47 on Chinese cabbage seedlings were analyzed in this study. After a 30-day hydroponic exposure to BDE-47 at different concentrations (25, 50, 75, and 100 µg·L−1), the fresh weight of Chinese cabbage seedlings was significantly decreased, whereas their root:shoot ratio was increased, indicating that BDE-47 inhibited the growth of the plant, especially the overground parts. The water content, chlorophyll content, and protein content of Chinese cabbage leaves also markedly decreased with the increase of the BDE-47 concentration. In addition, BDE-47 weakened the photosynthetic capacity of the leaves, which was supported by the decreased photosynthetic parameters [net photosynthetic rate (P n) and stomatal conductance (g S)]. Although the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in the leaves were enhanced after exposure to BDE-47, the increased malondialdehyde (MDA) content attested to the existence of membrane lipid peroxidation. The increased plasma membrane permeability and the decreased chlorophyll fluorescence parameters [the maximum quantum yield of PSII photochemistry at t = 0 (F v/F m), photosystem II (PSII) reaction centers (RCs) per cross section (CS) (RC/CS), absorption energy flux per CS (ABS/CS), trapped energy flux per CS (TR o/CS), electron transport flux per CS (ET o/CS), performance index on the absorption basis (PI abs), and driving force for photosynthesis (DF)] further proved that the plasma membrane and photosynthetic membrane were damaged by BDE-47. Our study demonstrated the phytotoxicities of BDE-47 to Chinese cabbage, which can provide valuable information for understanding the toxicity of BDE-47 on vegetables.

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Fang Yu, Zhiming Ni, Xingfeng Shao, Lina Yu, Hongxing Liu, Feng Xu and Hongfei Wang

To explore differences in sucrose metabolism between peach fruit subjected to chilling stress (5 °C) and nonchilling stress (10 °C), sucrose concentration as well as the activities and gene expression levels for enzymes associated with sucrose metabolism were compared. Fruits stored at 5 °C accumulated higher concentrations of H2O2 and developed severe chilling injury (CI) compared with fruit kept at 10 °C. Activities and gene expression levels for enzymes related to sucrose metabolism, such as acid invertase (AI), neutral invertase (NI), sucrose synthase (SS), and sucrose phosphate synthase (SPS) were higher in fruit stored at 5 °C than at 10 °C throughout or late in storage. A sharp increase in net sucrose cleavage activity dramatically decreased sucrose concentration and increased reducing sugars at 5 °C. The sucrose concentration at 10 °C increased over the first 21 days and then declined slightly, and was higher than in fruit at 5 °C throughout storage. The increase in net sucrose cleavage activity at 5 °C is contrary to the expectation that biochemical reactions ordinarily proceed more rapidly with increasing temperature. We conclude that chilling stress stimulates the activities and transcription levels of enzymes involved in sucrose metabolism, resulting in increased sucrose cleavage.