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    20 September 2022, Volume 42 Issue 5
    Orthotrichum tortifolium Lewinsky(Orthotrichaceae, Bryophyta), a Bhutan Moss New to China
    Qinghua WANG, Xiao’ao ZHENG, Yong JIANG, Yu JIA
    2022, 42(5):  721-725.  doi:10.7525/j.issn.1673-5102.2022.05.001
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    Orthotrichum tortifolium Lewinsky, an endemic to Bhutan, was recently first reported in China. Two specimens were collected by us in April, 2021 from Gongga Mountain Nature Reserve, Sichuan. The morphology was well consistent with the type of O. tortifolium. It was remarkably characterized by the flexuose to crisped leaves and the high basal membrane of endostome segments, which were rare in Orthotrichum. We provided detailed morphological description and photos here, and discussed its main differences from close species.

    Correction of Typographical Errors in the Protologue of 11 Taxon Names in Vascular Plants in China
    Yuxiu CHEN, Yun LIN, Shanshan HE
    2022, 42(5):  726-732.  doi:10.7525/j.issn.1673-5102.2022.05.002
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    According to Article 9.2 of International Code of Nomenclature for algae, fungi, and plants(Shenzhen Code), the typographical errors in the protologues of 11 taxon names in vascular plants in China were corrected, including Ostodes kuangii Y. T. Chang, Carpinus lancilimba Hu, Santiria yunnanensis Hu, Angiopteris caudipinna Ching, Astragalus longicalyx C. C. Ni & P. C. Li, Camellia vietnamensis T. C. Huang ex Hu, Onosma waddellii f. latifolium W. T. Wang, Viola microdonta C. C. Chang, Cyclosorus scaberulus Ching, Acer chunii subsp. dimorphophyllum Fang and Miscanthus lutarioriparius L. Liu ex S. L.Chen & Renvoize. The holotypes of these names were deposited at China National Herbarium(PE), Institute of Botany, Chinese Academy of Sciences.

    Systematic and Evolutionary
    Morphology and Taxonomic Significance of Pseudoparaphyllia in Brachytheciaceae (Bryophyta)
    Bojia WEI, Xuewen ZHANG, Youfang WANG, Jiancheng ZHAO, Min LI
    2022, 42(5):  733-740.  doi:10.7525/j.issn.1673-5102.2022.05.003
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    Brachytheciaceae is one of the most diverse and problematic taxonomic families in pleurocarpous mosses. In this paper, the morphology of pseudoparaphyllia of 594 specimens in Brachytheciaceae, which belonged to 19 genera and 44 species, were studied. Results were as follows: ①In Brachytheciaceae, pseudoparaphyllia was triangular to semicircular foliose. According to its apical characteristics, pseudoparaphyllia could be divided into two types: truncate or obtuse, and acuminate. ②Except for the genus Brachythecium, morphology of pseudoparaphyllia among the different species in the other 18 genera was basically the same. ③Morphology of pseudoparaphyllia had taxonomic significance in the genus level.

    Supplementary Description on Morphology and Cytology of Aster longipetiolatus C.C.Chang (Asteraceae)
    Jiawei XIAO, Yingjia WANG, Wenxuan HUANG, Weiping LI, Gongping KANG
    2022, 42(5):  741-745.  doi:10.7525/j.issn.1673-5102.2022.05.004
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    The morphological description of Aster longipetiolatus C.C.Chang was inadequate in the original literature when compared with the specimen which we collected from its type locality, and there was a lack of cytological reports. The detailed morphological and cytological description of A. longipetiolatus were carried out, and the new evidences of plant taxonomical revision would be provided. The results showed that: ①There were some newly-discovered characteristics for A. longipetiolatus. For example, the receptacle was convex, phyllaries 3- or 4-seriate, ray floret staminodes, lamina pilosulose and achenes 5-ribbed; ②Its cytology evidence revealed that the chromosome number was 2n=18 and the karyotype formulae was 2n=2x=16m+2sm, belonged to the type 1A.

    Anatomical Structure of Seed and Morphological Character at Different Germination Stages of Sinopodophyllum hexandrum
    Xiaolu CAO, Qiaozhu ZHAO, Hua XING, Mengfei LI
    2022, 42(5):  746-752.  doi:10.7525/j.issn.1673-5102.2022.05.005
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    To reveal whether the testa and endosperm are main factors restricting the seed germinationof Sinopodophyllum hexandrum, anatomical structure of seed in different germination stages(1, 7, 14, 21 and 28 d) of S. hexandrum were observed by using the tissue sectioning microscopy, and morphological characteristics of plants were investigated after the germination seeds sowing 7-210 d. The seed of S. hexandrum was composed of testa, endosperm and embryo. The testa contained episperm and endopleura, the episperm was compact and regular and it was composed of palisade cells and epidermal cells from outside to inside, and the endopleura was composed of 5-6 layers sponge cells. The endosperm accounted for most of the seed, included micropylar endosperm and outer endosperm. The embryo consisted of radicle, hypocotyl and cotyledons, surrounded by the compacted testa, multilayer-micropylar endosperm and outer endosperm. During the germination stage, the radicle and hypocotyl began to elongate from 1-7 d, two cotyledons separated from 7-14 d, the radicle broke through the micropylar endosperm and testa from 14-21 d, the radicle, hypocotyl and cotyledons continued to expand and elongate from 21-28 d. 3 function leaves and 5 adventitious roots were formed on average after seeding for 210 d. The compacted testa(physical dormancy) and multilayer endosperm(mechanical dormancy) were the two main factors that restricted seed germination of S. hexandrum.

    Plant Endemism in Northeast China
    Xiaojie CAO, Wei CAO, Yue ZHANG, Jia GUO, Yan GAO
    2022, 42(5):  753-761.  doi:10.7525/j.issn.1673-5102.2022.05.006
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    In order to better understand the origin of plant, phylogenetic differentiation and evolution process of flora, to promote the protection of endemic plants, endemic flora regions in Northeast China were systematically investigated respectively, including the species composition, family structure and genus structure, life-form composition, flora characteristics and geographical distribution. The results showed that:①There were 289 species of Chinese endemic plants, 109 species of endemic plants of Northeast China, 154 species of endemic plants of Changbai flora region, 16 species of endemic plants of Daxing’anling flora region, 10 species of endemic plants of Mongolia steppe flora region and 20 species of endemic plants of Northern China flora region. ②Among all kinds of endemic plants, herb were the most, and perennial herbs were the most dominant, while trees, shrubs and vines were the less. ③Chinese endemic plants were densely distributed in Daxing’anling Mountains and Changbai Mountains. ④The concentrated distribution areas of endemic plants of Northeast China were distributed along the main mountains in Northeast China, indicated that the endemic plants of Northeast China mainly originated from mountains. ⑤Changbai flora region had high level of endemism, showed the mature nature of the flora. Daxing’anling flora region, Mongolia steppe flora region and Northern China flora region had low level of endemism, showed the young nature of the flora. ⑥Endemic plants of Changbai flora region were densely distributed in Changbai Mountains. The endemic plants of Daxing’anling flora region were evenly distributed, and the endemic plants of Mongolia steppe flora region and the endemic plants of Northern China flora region were scattered.

    Genetic and Breeding
    Effect of Fruit-Expander on Cone Performance of Pinus sylvestris var. mongolica
    Yujin XU, Xihe WANG, Pingyang LI, Yulei LI, Dexiao LI, Yunyang WU, Quan SUN, Lili GE, Xiyang ZHAO
    2022, 42(5):  762-771.  doi:10.7525/j.issn.1673-5102.2022.05.007
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    Mongolian pine(Pinus sylvestris var. mongolica) is an important timber, landscaping and afforestation tree species in North China. In order to provide theoretical basis for high quality, high yields and stable yields of cones and putting forward scientific management strategies, the effect of fruit-expander treatment on cones and nuts characters of P. sylvestris var. mongolica was studied. P. sylvestris planted in Qinhe seed orchards, Wangqing County, Yanbian, Jilin Province were used as materials to compare and analyze the effects of traits, including cone yield, cone quality, seed yield and seed quality, treated with two fruit-expanders including chlorinated urea and α-sodium-naphthyl acetate respectively. The average seed traits were higher than those of the control, indicating that fruit-expander treatment could promote the seed growth of P. sylvestris var. mongolica. The cone weight, cone yield, seed long and seed width of P. sylvestris var.mongolica were all significant in different variation sources(P<0.001), indicating that the fruit-expander treatment had a significant effect on the cone quality and yield. The phenotypic variation coefficient of cone yield and seed weight reached to 28.54%, 33.88%, respectively. The cone traits were comprehensively evaluated for each treatment, and the optimal treatment was 10 mg·L-1 Chlorinated urea, and the real gains of cone length, cone width and cone weight were 11.81%, 4.65% and 24.35%, respectively. The optimal treatment according to the seed traits was 10 mg·L-1α-sodium-naphthynl acetate, and the real gains of seed yield per cone, seed weight, seed long, seed width, wing long and wing width were 21.32%, 24.15%, 0.24%, 9.16%, 0.33% and 2.16%, respectively. The results showed that chlorinated urea(10 mg·L-1) and α-sodium-naphthyl(10 mg·L-1) treatments could significantly improve the cone size and seed bearing of P. sylvestris var. mongolica.

    Comparison of Genetic Diversity Between Wild and ex-situ Conservation Populations of Cupressus chengiana
    Ermei CHANG, Jianfeng LIU, Yuening HUANG, Hongli LI, Bingyan SHAN, Zeping JIANG, Xiulian ZHAO
    2022, 42(5):  772-779.  doi:10.7525/j.issn.1673-5102.2022.05.008
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    To evaluate the genetic diversity level and provide theoretical basis of ex-situ conservation populations of Cupressus chengiana, the genetic diversity, genetic structure, and gene flow of ex-situ conservation populations and wild populations of C. chengiana were studied respectively. The principal component analysis(PCA), cluster analysis, construction of molecular evolutionary tree, genetic diversity and genetic structure of three wild populations and ex-situ conservation populations based on SNP locus were obtained respectively from Genotyping-by-Sequencing technology(GBS) sequencing. The results showed that high quality clean data 118 321 514 728 bp was obtained by GBS sequencing, and 1 947 047 tags were developed. After being screened, a total of 1 259 610 of high-quality SNP sites were retained. The valley value of cross validation error rate determined that the optimal number of clusters was 1. The phylogenetic tree indicated that most of individuals ex-situ conservation populations and wild populations were clustered together. Both of combined ex-situ conservation populations and wild populations of C. chengiana had higher genetic diversity, and their value of heterozygosity(Ho), expected heterozygosity(He), Shannon information index(Shi), inbreeding coefficient(Fis), and percentage of polymorphic loci(Pic) were 0.181 5-0.272 0, 0.223 2-0.300 3, 0.331 0-0.464 9, 0.178 0-0.246 5 and 0.272 2-0.309 2, respectively. Genetic diversity of ex-situ conservation cultivated populations was relatively higher than the wild populations of C. chengiana, the value of He and Shi was 0.300 3 and 0.464 9, respectively. The wild populationsof C. chengiana of Baiwan tunnel(BW)_vs_Songgang town(SA) had the greater genetic differentiation and the gene flow(Nm) were lower than other populations(Fst=0.091,Nm=2.496). The results indicated that genetic differentiation had not occurred among these C. chengiana populations, and the gene had exchanges frequently among the ex-situ conservation populations and wild populations of C. chengianaFst<0.05, Nm>4). There was a close genetic relationship between the ex-situ conservation individuals and the wild populations individuals, and there was no obvious clustering phenomenon. Thus, transplanting endangered plants was a better method in ex-situ conservation. This paper would provide a reference for the ex-situ conservation populations of wild populations C. chengiana and provided a theoretical reference for the conservation of other tree germplasm resources.

    Genetic Diversity Analysis from Different Geographic Populations of Linaria vulgaris var. sinensis by rpl32-trnL Fragment
    Yujie ZHANG, Pengxia SHI, Fan ZHAO, Xiaodan CHEN, Kun SUN, Hui ZHANG
    2022, 42(5):  780-789.  doi:10.7525/j.issn.1673-5102.2022.05.009
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    In order to investigate the population genetic characteristics of Linaria vulgaris, four different geographic populations containing 62 individuals were analyzed by using rpl32-trnL fragment of chloroplast DNA. The results showed that there were 15 haplotypes and 76 loci existing in 4 populations. The total genetic diversity of L. vulgaris reached a high level(Hd=0.878, π=0.003 88, K=2.994). The AMOVA analysis showed that the genetic variations were mainly maintained within the populations(51.49%), which was consistent with the genetic characteristics of self-incompatibility in L. vulgaris. The genetic differentiation among four geographic populations was significant(FST=0.466 14), while the gene flow between different geographic populations was rather low(Nm=0.29). The genetic differentiation between different populations was positively correlated with the geographical distance but no significance(R2=0.36, P>0.05). The results of neutrality test showed that the Heshui population(HS) might experience some bottleneck events during its evolution (Fu and Li’s D=-2.450 49,P<0.05), and the evolutionary feature of the other populations was consistent with the neutral hypothesis. This study revealed that the reproductive strategies, geographic isolation, and habitat disturbance were not only responsible for the rather high level of genetic diversity in the populations distributed in the northern area of China as well as their genetic structure characteristics, but also it might provide a scientific basis for the further study of conservation genetics in L. vulgaris.

    Molecular biology
    Identification of Glutathione S-transferase Gene Family in Sugar Beet and the Response to Cadmium Stress
    Xin’ai ZHONG, Shiqi MENG, Wanting Zhou, Qi YAO, Qiong ZHANG, Wang XING, Dali LIU
    2022, 42(5):  790-801.  doi:10.7525/j.issn.1673-5102.2022.05.010
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    To clarify the potential function of Glutathione-S-transferase gene(GST) family in sugar beet(Beta vulgaris) growth and development and abiotic stress, the sugar beet BvGSTs was identified, and the physical and chemical properties, evolutionary relationship, homeopathic elements, chromosome location and transcriptional expression characteristics under cadmium stress were comprehensively analyzed, respectively. The results showed that there were 52 members of BvGSTs in sugar beet genome, which were distributed in seven subfamilies; The molecular weight of the members ranged from 17-28 kDa, and the theoretical isoelectric point(pI) was between 5.16-6.77, and most of the members were located in cytoplasm. There were nine motifs found in BvGSTs, among which motif eight was unique to Phi subfamily. Among all the members, 30 BvGSTs were distributed in eight chromosomes respectively, with four tandem repeats. According to cis-acting element analysis, BvGSTs might be involved in a variety of biotic and abiotic stress responses. Transcriptome analysis showed that all of 52 BvGSTs were related to response of sugar beet to cadmium stress. The expression of most members of Tau subfamily under ground parts was positively regulated by cadmium stress; Phi subfamily above ground parts was significantly induced by cadmium stress, and the expression of Tau family was inhibited. The result of qRT-PCR showed that the transcripts of the four differentially expressed BvGSTs were regulated by cadmium stress, which was consistent with the results of transcriptome sequencing. These results would play a foundamental role in further study on the biological function of sugar beet glutathione S-transferase under cadmium stress.

    Structural and Gene Expression Analysis of a DUF1262 Domain Protein in Latex from Hevea brasiliensis
    Longjun DAI, Mingyang LIU, Jianghua YANG, Kai ZHOU, Bingbing GUO, Hong YANG, Lifeng WANG
    2022, 42(5):  802-810.  doi:10.7525/j.issn.1673-5102.2022.05.011
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    To reveal the structure and function of natural rubber biosynthase interacting proteins in natural rubber biosynthesis, the total protein of latex rubber particles of Hevea brasiliensis was used as material, and an unknown functional protein containing DUF1262 domain was captured by immunoprecipitation assay using an antibody targeting CPT, a key enzyme for natural rubber biosynthesis. Bioinformatics analysis showed that there were 50 gene sequences encoding DUF1262 domain proteins in the rubber tree genome, and protein interaction network analysis suggested that DUF1262 domain proteins might be involved in signal transduction and transcriptional regulation. Fluorescence quantitative PCR results showed that the transcription of the gene encoding this protein was widely distributed in roots, leaves, flowers, branches, latex and other tissues respectively, andthe expression was low in latex and but high in bark. Salicylic acid, abscisic acid, hydrogen peroxide and drought treatment could enhance the transcription level of this gene in leaves respectively. This study demonstrated that the DUF1262 protein was involved in stress response and other physiological processes of rubber tree, providing new clues for revealing the regulation mechanism of latex biosynthesis.

    Cloning and Functional Analysis of Heat Shock Protein HbHSP90.8-1 from Hevea brasiliensis Müll. Arg.
    Mingyang LIU, Huaxing XIAO, Lifeng WANG, Xiaoxu LIANG, Yu ZHANG, Meng WANG
    2022, 42(5):  811-820.  doi:10.7525/j.issn.1673-5102.2022.05.012
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    Powdery mildew is one of the major leaf diseases in growth of rubber tree. Heat shock protein 90(HSP90) molecular chaperone plays an important role in plant stress resistance. To study the structure of the rubber tree HSP90 family members and their function in powdery mildew resistance, HbHSP90.8-1 was cloned from rubber tree variety Reyan73397 leaf using PCR technology and its structure and function were analyzed by bioinformatics method. The results showed that the cDNA sequence of HbHSP90.8-1 was 2 844 bp in full length, 2 454 bp open reading frame(ORF) and encoded 817 amino acids. HbHSP90.8-1 encoded a stable hydrophilic protein with signal peptide, no transmembrane structure and predicted positioning on endoplasmic reticulum, contained HSP90 superfamily and HATPase superfamily domains. Phylogenetic analysis showed that HbHSP90.8-1 was closely related to MeHSP90, and classified into Jatropha JcHSP90 and Castor RcHSP90. The qRT-PCR analysis showed that HbHSP90.8-1 was expressed in different tissues of rubber tree, and its expression level was the highest in latex. The expression of HbHSP90.8-1 was significantly expressed and up-regulated in leaves treated with Oidium heveae infection, H2O2, ABA and ETH respectively. However, under the treatment of MeJA and SA hormone, the expression of HbHSP90.8-1 showed a downward trend. The study suggested that HbHSP90.8-1 related to the powdery mildew resistance and the hormone signal transduction pathway related to plant disease resistance, and provided a technical guidance for the subsequent study on the molecular regulation mechanism of HbHSP90.8-1 in the powdery mildew resistance in rubber tree.

    Cloning and Functional Analysis of StNPR4 gene in Solanum tuberosum
    Denggao LI, Rui LIN, Qinghui MU, Na ZHOU, Yanru ZHANG, Wei BAI
    2022, 42(5):  821-829.  doi:10.7525/j.issn.1673-5102.2022.05.013
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    In order to clarify the function of StNPR4 in potato, the CDS and promoter sequence of StNPR4 were cloned and bioinformatics analysis was conducted. Tissue expression pattern was performed by qRT-PCR, and a binary expression vector of StNPR4 driven by its own promoter was constructed and transformed into potato. Transgenic plants were used to examine the responses to salicylic acid, Phytophthora infestans and high salt stress respectively. The results showed that StNPR4 had typical functional domains in NPR1 family with cis-acting elements on the promoter in response to biotic and abiotic stresses. The StNPR4 expression level was highest in leaves StNPR4 was inductively expressed by SA and the induced expression was higher in transgenic plants than in the control. StNPR4 transgenic potato plants showed enhanced resistance to Ph. infestans and higher rooting rate under high salt stress. This indicated that StNPR4 played an important role not only in biotic stress but also in abiotic stress of potato.

    Identification and Expression Pattern Analysis of TkAPC10 in Taraxacum kok-saghyz Rodin
    Bi QIN, Xiaoxiao WANG, Yushuang YANG, Qiuhai NIE, Qiuhui CHEN, Shizhong LIU
    2022, 42(5):  830-839.  doi:10.7525/j.issn.1673-5102.2022.05.014
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    APC/C is an E3 ubiquitin-ligase complex that is involved in regulating cell-cycle progression. In order to reveal the function of the APC/C complex in Taraxacum kok-saghyz Rodin(TKS), TkAPC10 was identified, and its expression pattern was analyzed to preliminarily determine its function in this study. The ORF of TkAPC10 was 579 bp, encoding 192 amino acids. Its genomic DNA sequence was 1 092 bp, contained 6 exons and 5 introns. Genome analysis showed that TkAPC10 was a single copy in TKS genome. In addition to the TATA-box and CAAT-box enhancer elements, TkAPC10 promoter contained cis-acting elements related to ABA, JA, light, and stress response. Phylogenetic analysis suggested that APC10 from different species showed high homology. The similarity between TKAPC10 and lettuce LsAPC10 was up to 99%, and the similarity to APC10 from other Compositae plants was also over 95%. The expression pattern of TKAPC10 was analyzed by qRT-PCR, and the results indicated that TKAPC10 expression in the tissues with vigorous cell division(such as flowers, leaves and roots) was significantly higher than that with relatively slower cell division (such as pedicel). Under exogenous ABA treatment, the transcript level of TKAPC10 was obviously decreased, while it was significantly increased after MeJA and ET treatments. Under PEG6000 and mannitol treatments, the expression of TKAPC10 was obviously decreased, but its expression was markedly induced by salt stress treatment. The results in this study suggested that TKAPC10 was involved in the regulation of cell division, hormone signaling and abiotic stress response in TKS.

    Differential Expression of miRNA396 and CKX Genes in Eucalyptus urophylla × E. grandis Callus Induced by Different Concentrations of Novel Mitogen PBU
    Yamei LIU, Yupeng WU, Limei LI, Min SU, Zhu YU, Lejun OUYANG
    2022, 42(5):  840-847.  doi:10.7525/j.issn.1673-5102.2022.05.015
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    Eucalyptus plays a great role in ecological environment and chemical industry. As a hybrid of Eucalyptus urophylla and E. grandisE.urophylla × E.grandis has good heterosis and is more synthetical than other Eucalyptus species. In order to meet the requirements of cultivating superior Eucalyptus cultivars with different uses by genetic engineering technology, it is of great significance to establish an efficient in vitro regeneration system of E.urophylla × E.grandis and study its callus differentiation mechanism. The growth and differentiation of E.urophylla × E.grandis callus is affected by its endogenous phytohormones and miRNA396 is a small RNA regulating the growth and development of plant leaves and roots, which is related to the biosynthesis of cytokinin. CKX is responsible for regulating cytokinin oxidase gene. In order to explore the regulatory effects of miRNA396 and CKX genes on the growth and development of E.urophylla × E.grandis callus, in this study, the genome of E.urophylla × E.grandis was used as a template for PCR amplification and sequencing to analyse the miRNA396 sequence of E. urophylla × E.grandis gene. The cDNA reverse transcribed by RNA from E.urophylla × E.grandis callus cultured on media with different PBU cytokinin concentrations was used as a template by fluorescence quantitative PCR and the expression differences of miRNA396 and CKX in E.urophylla × E.grandis callus treated with different PBU concentrations were determined. The results showed that compared with the E.urophylla × E.grandis callus treated with 0.5 mg·L-1 PBU, the expression of miRNA396, CKXACKXB and CKXF in E.urophylla × E.grandis callus treated with 1 mg·L-1 PBU was significantly decreased, and the difference reached a very significant level. CKXCCKXD and CKXE were up-regulated, but only the relative expression of CKXC reached a very significant level. The expression levels of miRNA396A, CKXDCKXE and CKXF in E.urophylla × E.grandis callus treated with 2 mg·L-1 PBU were down-regulated, and the difference reached a very significant level. The rest of gene expression levels were up-regulated. The relative expression levels of CKXACKXB and CKXC reached a very significant level. This study preliminarily established the regulation and expression differences of miRNA396 and CKX gene in the callus of E.urophylla × E.grandis, laid a foundation for the subsequent analysis of the miRNA regulation network of E.urophylla × E.grandis, and provided a certain reference for the establishment of an efficient regeneration system of E.urophylla × E.grandis.

    Gene Cloning and Specific Tissue Expression of UlGLK from Ulmus laevis
    Xiangdong BAI, Chengrui GU, Xiaoyue ZHANG, Jing JIANG
    2022, 42(5):  848-854.  doi:10.7525/j.issn.1673-5102.2022.05.016
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    GLK transcription factor is a member of GARP transcription factor superfamily, which plays a role in regulating plant chloroplast development, biological and abiotic stress, senescence and hormone signal transduction. GLK had been successfully used in leaf color improvement of Betula platyphylla and Populus alba × P. berolinensis, etc. In order to explore the function of UlGLK gene in Ulmus laevis and provide reference for molecular design and breeding of U. laevis. UlGLK was cloned from U. laevis and sequenced and the internal reference genes were screened for qRT-PCR analysis and the tissue expression characteristics of UlGLK gene was analyzed. The results showed that the ORF of UlGLK gene was 1 353 bp in length, encoded 450 amino acids, which had high homology with cannabis sativa and Morus notabilis. Based on the expression levels of UbiquitinActin and Tubulin genes in U. laevis root, stem and leaf respectively, ubiquitin gene was identified as an internal reference for qRT PCR analysis. Analysis of tissue expression characteristics of UlGLK showed that the expression level of UlGLK was the highest in leaf, but lower in root and stem. The results provided a reference for further molecular breeding of U. laevis.

    Gene Cloning and Interaction Proteins Screening of DcPMK in Dioscorea composite
    Hongpeng WANG, Yidan LI, Yao WANG, Xiaoyu TAN, Chengbin CHEN, Lipeng ZHANG
    2022, 42(5):  855-865.  doi:10.7525/j.issn.1673-5102.2022.05.017
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    To explore the synthesis path of terpenoid in Dioscorea composite and to provide theoretical reference for studying, D. composite was used as material, DcPMK gene was cloned by RT-PCR and the bioinformatic analysis, tissue-specific expression analysis and expression analysis after salicylic acid (SA) treatment were carried out respectively, and the bait vector of DcPMK was constructed to screen interacting proteins from Arabidopsis yeast library. The results showed that the open reading frame (ORF) of DcPMK was 1 536 bp (GenBank accession No. MZ171241), which encoded 511 amino acids. By multiple sequence alignment, the results showed that similarity between DcPMK and PMK amino acid sequence of other plants was as high as 88.85%, DcPMK obtained 1 ATP binding site Gly-X-Gly-XX-Ala. The result of phylogenetic tree showed that DcPMK was closely related to monocotyledon such as Phonenix dactylifera.HPLC analysis showed that the content of diosgenin was the highest in rhizome, qRT-PCR analysis indicated that DcPMK was constitutively expressed among flower, leaf, stem, rhizome and root, with the highest levels in old stem and the lowest in rhizome. After SA treatment, the changes of diosgenin content were in parallel with the expression levels of DcPMK, the diosgenin content was improved as DcPMK up-regulated expression in leaf. Yeast two-hybrid results showed that DcPMK was no autoactivation and cell toxicity, then 27 significant interactional genes were screened, which were growth and development-related genes AtKCR1(AT1G67730), AtRPS9M(AT3G49080), AtASY4(AT2G33793), abiotic and biotic stress-related genes AtVDAC2(AT5G67500), AtVDAC3(AT5G15090), AtRH8(AT4G00660), anthocyanin accumulation-related gene AtPHR2(AT2G47590). The results illustrated that DcPMK could participate in the synthesis of terpenoids in D. composite, and was widely involved ingrowth and stress response pathways through protein-protein interaction.

    Physiology and Ecology
    Allelopathic Effects on Seed Germination and Seedling Growth of Brassica pekinensi, Caused by Water Extracts of Branches and Leaves from Davidia involucrata and Bothrocaryum controversum
    Huan WANG, Yunfei XU, Yibo LIU, Qinsong LIU, Wenjuan XU, Yun LONG, Xiao XU
    2022, 42(5):  866-875.  doi:10.7525/j.issn.1673-5102.2022.05.018
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    In order to identify whether Davidia involucrata and its associated tree species Bothrocaryum controversum affect the seed germination and seedling growth of other plants through allelopathic effects to maintain their stable associated relationship, the branches and leaves of wild D. involucrata and B. controversum in Pingwu County were used as the material source of the water extract, and the seeds of Brassica pekinensi were explored the effects of water extracts of the branches or leaves of the two species or their mixture at five concentrations(0, 0.025, 0.050, 0.075, 0.100 g·mL-1) on the seed germination and seedling growth of B. pekinensi respectively. The results showed:①Water extracts of branches of D. involucrata and Bothrocaryum controversum had little effect on the germination of Brassila pekinensi seeds, but seedlings growth was significantly promoted and increased with increasing concentration(P<0.05). ②The water extracts of D. involucrata and Bothrocaryum controversum had significant inhibitory effects on seed germination of Brassila pekinensiP<0.05), but their effects on seedling growth were different. The former showed the phenomenon of “low promotion and high inhibition” with the increase of concentration, while the latter showed the characteristic of “inhibition increase”. ③The water extracts from different sources of branches-leaves mixtures had significant inhibitory effect on seed germination of Brassila pekinensiP<0.05), and showed the characteristics of “low promotion and high inhibition” with the increase of concentration. The results demonstrated that the allelopathic effects of water extracts from branches and leaves of D. involucrata and Bothrocaryum controversum on seed germination and seedling growth of the same recipient plant were different, and different species, parts and concentrations could lead to different allelopathic intensity and direction. This phenomenon might be a long-term plant evolution and maintained the stable interspecific relationships.

    The Correlation Between Leaf Phenotype Diversity and Total Flavonoids Content of Overground Part of Tetrastigma hemsleyanum Diels & Gilg
    Mengqiao GUO, Xuanyu CHEN, Senrong HONG, Jiao LI, Jie FAN, Xinyu CHENG
    2022, 42(5):  876-885.  doi:10.7525/j.issn.1673-5102.2022.05.019
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    To reveal the variation of leaf phenotypic diversity of Tetrastigma hemsleyanum Diels & Gilg from different origin and its correlation with the total flavonoids content of overground part. In this paper, 49 germplasm resources(abbreviation resources) of T. hemsleyanum were selected as materials, and 12 characters of length and 7 proportion characters of leaf phenotype were compared. The phenotypic characters of leaf and their relationship with origin location and total flavonoids content were statistically analyzed by correlation analysis, factor analysis and cluster analysis. The results showed that there were significant differences in leaf length characters among resources, and the leaf characters of petiolule and lateral leaflet were of high reference value in the identification of T. hemsleyanum resources; There was correlation between leaf length characters and proportion characters, which showed that leaflet was wider when petiole grew, and leaflet was longer and narrower while leaflet elongated; Some leaf shape characters, such as central leaflet length and lateral petiolule length, were significantly(P<0.05) or extremely significantly(P<0.01) correlated with latitude, longitude or altitude of origin location, which indicated that these characters could be used as preliminary basis for geographical location estimation of origin; The cluster analysis based on 3 common factors of leaf shape showed that the leaf shape of 49 resources of T. hemsleyanum could be divided into five types, and the number of leaf type in each origin(municipalities) showed a radially decreasing trend from Jiangxi to the surrounding area; There was no significant correlation between the total flavonoids content of the overground parts and the geographical location of origin, but total flavonoids content was significantly negatively correlated(P<0.01) with the common factor of leaf blade characters(common factor 1), and significantly positively correlated(P<0.01) with the common factor of petiole length characters(common factor 2). The results improved the morphological data of T. hemsleyanum and provided the morphological basis for the breeding of excellent varieties of T. hemsleyanum.

    AM Fungi Inoculation on Root Morphology and Nutrient Loading of Clematis fruticosa Seedlings under Simulated Atmospheric Nitrogen Deposition
    Zhaoyi LI, Longfei HAO, Tingyan LIU, Yanhong HE, You ZHANG, Shulan BAI, Xinyu YANG
    2022, 42(5):  886-895.  doi:10.7525/j.issn.1673-5102.2022.05.020
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    In order to investigate the stress response of root morphology and nutrient loading of Clematis fruticose mycorrhizal seedlings to nitrogen deposition, 1-year-old C. fruticose mycorrhizal and non-mycorrhizal seedlings(-M) were used as materials. The inoculation treatments included single-inoculation(Rhizophagus intraradices, +R; Funneliformis mosseae, +F), mixed-inoculation(AMF agents 1∶1 mixture, +RF). Four nitrogen deposition treatment were set, including no nitrogen(CK, 0 g·m-2·a-1), low nitrogen(LN, 3 g·m-2·a-1), medium nitrogen(MN, 6 g·m-2·a-1), high nitrogen(HN, 9 g·m-2·a-1). The fine root morphology with diameter of ≤0.5 mm(including total root length, total surface area, total volume and tips), mycorrhizal colonization rate, soil spore density and all parts nutrient loading of a plant(including carbon, nitrogen and phosphorus content) were determined under inoculation and nitrogen deposition treatments. The results indicated that: ①Under +R and +RF treatments, the seedlings mycorrhizal colonization rate and soil spore density were the maximum in LN treatment, and the mycorrhizal colonization rate in LN treatment was significantly higher than that in HN treatment. Under +F treatment, the mycorrhizal colonization rate was no significant difference between the different nitrogen deposition treatments. ②Under 0N treatment, the fine root morphology with diameter of ≤0.5 mm in +F and +R treatments, including total root length, total surface area, total volume and tips, were higher than that in -M treatment respectively. However, the above root morphological indicators of C. fruticose seedlings were decreased with the increase of nitrogen deposition under +F and +R treatment. ③Under +F and +R treatments, the nutrient loading of seedlings increased with the increasing of nitrogen deposition content. The carbon, nitrogen and phosphorus(C,N and P) contents of mycorrhizal seedlings were significantly higher than that of non-mycorrhizal seedlings, and the seedlings C, N and P contents were the maximum under +F treatment. ④There was a positive correlation between fine root morphology with diameter of ≤0.5 mm and nutrient content of seedlings. Thus, inoculation treatment changed the response of fine root morphology of C. fruticose seedlings to nitrogen deposition. The F. mosseae effectively enhanced the seedlings adaptability to nitrogen deposition, and that improved the seedling nutrient loading under high nitrogen deposition.

    Comparative Analysis of Nutrient Elements of Eight Herbaceous Plants in Desert Area
    Weiwei ZHUANG, Mingming WANG
    2022, 42(5):  896-909.  doi:10.7525/j.issn.1673-5102.2022.05.021
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    In order to better understand the quantitative characteristics of nutrient elements in desert plants and the interaction among elements, the differences of plant element contents in different growing stages and dune areas were revealed. Eight dominant herbs(four kinds of annual plants, four kinds of ephemeral plant) in Gurbantunggut desert were used to collect plants from different growth period(vigorous period, wither period) and different sand dune areas(slopes, hinterland), and 10 kinds of nutrient elements(C, N, P, K, Na, Mg, Al, Mn, Cu and Zn) in whole plant were measured respectively. The results showed that: (1)There were significant differences in element contents of 8 species, which reflected the characteristics of selective absorption of elements due to different genetic characteristics. The contents were as follows: C(230.19-401.82 mg·g-1) and N(11.31-18.85 mg·g-1), P(0.95-2.08 mg·g-1), K(16.12-29.79 mg·g-1), Na(0.88-3.31 mg·g-1), Mg(3.38-5.31 mg·g-1), Al(0.33-1.99 mg·g-1), Mn(51.35-105.32 mg·kg-1), Cu(4.14-6.38 mg·kg-1), Zn (11.64-21.43 mg·kg-1). The distribution pattern of element contents of different species was similar, and the order of element contents was: C>N>K>Mg>Na>P>Al>Mn>Zn>Cu, typical characteristics were rich in K but poor in Cu and Zn, belonging to N restrictive plants, demonstrated consistent impact of environmental elements for all kinds of plants. (2)Element contents were significantly affected by species, growing period and dune location. The Na contents of four species of Chenopodiaceae were significantly higher than those of four ephemeral plants. Compared with the flourishing stage, the contents of C, N, P, Mg, Mn, Cu and Zn of the eight plants were generally decreased in the withered stage, while the contents of Na, K increased by different degrees, and the K content of Suaeda glauca and Agriophyllum sqarrosum significantly increased. The content of plant elements in the hinterland of sand dune was generally higher than that of the same species on the slope due to the influence of soil moisture and nutrient. (3)Correlation analysis showed that: C was positively correlated with P, N with Na, Mg, Cu, Zn, K with P, Al, Na with Mg, Mn with Na, Mg, Cu, Zn with Cu(P≤0.001) respectively, but K was negatively correlated with Mn, Cu and Zn(P≤0.001). In conclusion, the characteristics of element contents of desert plants reflected their adaptability to climate and soil in arid and semi-arid areas.

    Ecological Stoichiometric Characteristics of C, N and P and Their Relationship with Soil Factors from Different Organs of the Halophytic Chenopodiaceae Plants in Hulunbuir
    Yuhang SU, Xiaoqian SONG, Jingwen ZHENG, Zhonghua ZHANG, Zhonghua TANG
    2022, 42(5):  910-920.  doi:10.7525/j.issn.1673-5102.2022.05.022
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    The lack of available nutrients is one of the main factors limiting the biomass of saline plants, the largest group in the saline environment is Chenopodiaceae plants, and the overall nutritional strategy of Chenopodiaceae plants is of great significance to the breeding in saline-alkali soil and agricultural development. four typical Chenopodiaceae plants in Hulunbuir were used as materials, including Suaeda glaucaChenopodium acuminatumSalsola tragus and Bassia dasyphylla. In order to reveal the C, N, P stoichiometric characteristics of Chenopodiaceae plants and the relationship with soil factors, the ecological stoichiometric characteristics of carbon(C), nitrogen(N) and phosphorus(P) in different organs were analyzed respectively. The results showed: ①Both the stem N/P and leaf N/P of Chenopodiaceae plants were greater than 16, and the root N/P of Chenopodiaceae plants was less than 14; The content of C and N in each organ had a significant correlation, root C content>stem C content>leaf C content, The N content of each organ showed the order of leaf N content>stem N content>root N content, it showed that the N element had a good transfer efficiency from roots, stems to leaves. ②The content variability of C, N and P in various organs was compared, and P had the largest coefficient of variation. There was a significant positive correlation between leaf P and leaf N, stem P content also had a significant positive correlation with leaf N, root N content, and root P content had a significant negative correlation with leaf N and root N content. It indicated that the N and P elements had a strong coordination relationship in leaves and roots. ③The RDA ranking indicated that Soil P was the main factor affecting plant leaf stoichiometry, soil K was the main factor of stem stoichiometry variation, and soil N was the main factor of root stoichiometry variation. This study found that Chenopodiaceae plants promoted the accumulation the coordination of N, P in leaf, and reduced the impact on soil P and N restrictions, and showed great significance to saline-alkaline soil management.