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    20 May 2022, Volume 42 Issue 3
    Flora of Lycophytes and Ferns in Northeast China
    Yan LI, Wei CAO, Huan HE, Yue ZHANG, Daoyan SU, Wei CHEN
    2022, 42(3):  321-328.  doi:10.7525/j.issn.1673-5102.2022.03.001
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    Northeast China is the richest area of lycophytes and ferns in northern China, and systematic study on its flora is the basis and premise of regional lycophyte and fern resource management. In this study, based on literature analysis and specimen review, the flora and characteristics of lycophytes and ferns in Northeast China were explored. The results showed that: ①There were 141 taxa(species and intraspecific taxa), which belonged to 48 genera of 19 families. ②The flora of lycophytes and fern in Northeast China was temperate, with ancient tropical origins and cold zone components, which indicated the complexity of its geographical composition. ③Predominant families and genera were obvious: the number of species in seven dominant families accounted for 63.83% of the total number of species, and six dominant genera for 46.10%. There were also many monotypic genera and oligotypic genera with two or three species, accounting for 83.33% of the total number of genera, but 47.52% of the total number of species. ④Northeast China was an important distribution area of Equisetaceae in China. ⑤A total of 30 Chinese endemic species of lycophytes and ferns were distributed in Northeast China, of which four species were endemic to Northeast China(Huperzia asiaticaAthyrium yokoscense var. kirismaenseDeparia pycnosora var. longidens and Parathelypteris changbaishanensis). This study provided a necessary supplement to the study of flora in Northeast China, and had important theoretical and practical significance.

    Correction of Typographical Errors in the Protologue of Nine Taxa (Sympetalae) in China
    Yun LIN, Qian SUN, Yali CHEN, Xuan JING
    2022, 42(3):  329-340.  doi:10.7525/j.issn.1673-5102.2022.03.002
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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 nine taxa (Sympetalae) in China were corrected respectively, including Limonium drepanostachyum Ik.-Gal. ssp. callianthum T. X. Peng, Phlomis pararotata Sun ex C. H. Hu, Salvia chunganensis C. Y. Wu & Y. C. Huang, Scutellaria pekinensis Maxim. var. grandiflora C. Y. Wu & H. W. Li, Lindernia macrobotrys P. C. Tsoong, Pedicularis minutilabris P. C. Tsoong, Chirita shuii Z. Y. Li, Codonopsis levicalyx L. D. Shen and Jurinea flaccida C. Shih.

    Systematic and Evolutionary
    Application of Cuticular Micromorphology of the Cuticle of Pinus Needles in Taxonomy
    Jun LI, Yaping DUAN, Xiuzhen CAI, Ting WANG, Baihan PAN
    2022, 42(3):  341-351.  doi:10.7525/j.issn.1673-5102.2022.03.003
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    The cuticle is an impermeable fatty layer on the surface of the epidermal cell wall, and closely combined with epidermal cells, the micromorphological characteristics such as the morphology and arrangement of plant epidermal cells, as well as the morphological structure of the stomatal apparatus, can be reflected in the cuticle. In this paper, the cuticular micromorphology of needles of 12 species of Pinus was observed and compared by optical microscope and scanning electron microscope, and 20 characters were described in detail, of which 12 characters came from the inner surface of the cuticles and eight characters from the outer surface of the cuticles respectively. The results showed that these characters had taxonomic importance for the intra-genus classification and the identification of similar species: ①Micromorphological features of the cuticle such as the length of epidermal cells, the length of bristle, the undulating degree of the outer surface of the cuticle, the outline of epidermal cells, the presence or absence of stomatal plug and needle-like substances had their specificities, which could be used as a basis for the classification of Pinus at the section level. The micromorphological features of cuticles did not support the view of merging P. Section Cembra and P. Section Parrya into P. Section Quinquefolius, nor did it support the view of dividing P. Section Pinus into P. Section Pinus and P. Section Trifolius. ②The cuticle micromorphological characteristics of the needles of P. bungeana were partly the same as P. Section Cembra, partly similar to P. Section Pinus, and partly different from other species of Pinus, which could provide a new argument for the establishment of P. Subgenus Parrya. ③Under scanning electron microscope, the anticlinal wall pattern of epidermal cells, the existence or nonexistence of stomatal plug and the stomatal shape of the outer surface could provide interspecific delimitation basis for morphologically similar species of P. taeda and P. elliottii.

    Effect of Brefeldin A on the Development of Fern Eggs
    Yandong GUO, Jianguo CAO, Yifeng ZHOU, Xiling DAI, Quanxi WANG, Mingxia ZHANG
    2022, 42(3):  358-363.  doi:10.7525/j.issn.1673-5102.2022.03.005
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    Ceratopteris thalictroides L. Brongn. was treated with different concentrations of brefeldin A(BFA) for 10, 20, 30 h, respectively and the resin sections were observed by optical microscope and electron microscope. On this basis, the periodic acid schiff reaction(PAS) and Sudan black B reaction were carried out. The results showed that in the control, the egg of C. thalictroides in the middle stage of development was larger, the nucleolus was round, and the organelles such as Golgi body and endoplasmic reticulum were complete. The fertilization pore between egg and ventral canal cell was clearly visible and there was only a small amount of vesicular secretion in separation cavity between them. Egg membrane was thick with distinct layers. However, under the treatments, the fertilization pore and egg membrane were not typical, which were composed of a large number of vesicles with different sizes. There were more mitochondria and a few Golgi body under fertilization pore; membrane organelles such as Golgi body and endoplasmic reticulum, etc. were broken. A large number of osmiophilic vesicles were distributed above nucleus and below egg membrane and the separation cavity was filled with flocculent substances. Tissue localization showed that separation cavity of the control was empty, while those of treatments were full of polysaccharides. In conclusion, BFA could damage membrane organelles such as endoplasmic reticulum and Golgi body, etc., affected the normal function of secretory system first and then the formation of egg membrane and fertilization pore. The results of this study lay a foundation for further study on the mechanism of fertilization pore formation and the reproductive biology of ferns.

    Nectary Structure of Selfing and Outcrossing Speciesin Roscoea
    Wenjing WANG, Hongfan CHEN, Guojun SHAO, Hong LIAO, Jianli ZHAO, Qingjun LI
    2022, 42(3):  364-372.  doi:10.7525/j.issn.1673-5102.2022.03.006
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    Being the key floral structure of the reciprocal relationship between flowering plants and pollinators, the analysis of the differences of nectary structure among species with different reproductive characteristics could provide theoretical basis for understanding the evolution of reproductive characteristics of flowering plants. Two contrary species, Roscoea cautleoides with highly specialized outcrossing pollination system and R.schneideriana with autonomous selfing, were used to explore the biological importance of nectary. Nectary size and volume were recorded in the field and nectary location, external form and internal structure were observed by paraffin section staining method. The results showed that: both species had structural nectary. From the external morphology, R. cautleoides nectary was developed and secreted higher concentration sugar, while R. schneideriana nectary was wilting and had no nectar secretion. From the internal structure comparison, R. cautleoides nectary was specialized and the division of structure was clear, while R. schneideriana nectary structure appeared a degree of degradation. These results provid an important basis for revealing the differences of floral characteristics, pollination mechanism and breeding system, as well as the mechanism of nectar secretion between outcrossing and selfing species in Roscoea. It also lays a scientific foundation for further exploring the coevolutionary relationship between plants and pollinators, and understanding the origin and maintenance mechanism of nectary diversity.

    Genetic and Breeding
    Genetic Structure Analysis of Picea crassifolia Based on Genome-wide SNP Molecular Markers
    Hongbin ZHANG, Dong LÜ, Ming ZHAO, Hu ZHAO, Xingpeng ZHAO, Wei LI
    2022, 42(3):  373-382.  doi:10.7525/j.issn.1673-5102.2022.03.007
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    A total of 106 clones of Picea crassifolia in Longqu P. crassifolia Clone Seed Orchard in Zhangye were used as materials, and the genomic DNA of P. crassifolia was extracted by improved CTAB method, the SLAF library was constructed and sequenced with high flux, and then the SLAF sequencing data were analyzed and SNP sites were screened, and the clustering of samples was obtained based on adjacency analysis respectively. The results showed that the efficiency of two terminal alignment was 95.33%, indicated that the SLAF database was successfully established. The Q30 of the sequence measured and the sequencing quality was high, but the error rate of base sequencing was low. A total of 4 058 883 SLAF tags were developed, and the average sequencing depth of the tags was 21.21×. A total of 12 275 765 P. crassifolia SNP markers were developed, and the number of SNPs in each P. crassifolia sample ranged from 1 890 934-4 487 841. Using the developed high quality SNP markers of P. crassifolia, 106 phylogenetic trees of P. crassifolia were constructed. It was found that P. crassifolia from different provenances were evenly distributed in each group, and P. crassifolia from different provenances mostly were clustered into one class. By SNP markers and principal component analysis, these clones were more likely to be derived from the same ancestor, indicated that the genetic relationship between clones was similar. It provids basic data for the analysis of genetic diversity and the construction of genetic map in the future, and also provids a basis for the elite seeds selection in P. crassifolia primary seed orchard, and lays a foundation for the construction of high generation seed orchard.

    Genetic Variation and Joint Selection of Growth and Wood Traits in Half-sib Families of Larix olgensis
    Xueyan ZHOU, Biying WANG, Xuefeng HAO, Xinguo HU, Jiangtao WU, Kai LANG, Qinbo HU, Xiyang ZHAO
    2022, 42(3):  383-393.  doi:10.7525/j.issn.1673-5102.2022.03.008
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    As the important timber tree in Northeast China,comprehensive selection based on growth traits and wood traits was essential for Larix olgensis. A total of 49 half-sib families of L. olgensis at 32-year-old from Wangqing Forestry Bureau, in Yanbian Korean Autonomous Region of Jilin Province was selected as materials. Nine growth traits(tree height, ground diameter, diameter of breast height, diameter of stem at 3 m height, volume, taperingness, crown width, branch angle and stem straightness degree) and four wood traits(wood density, tracheid length, tracheid width and tracheid length-width ratio) were measured and analyzed respectively. The difference of all traits among different sources of variation reached extremely significant levels(P<0.01). The heritability of all traits was high(0.51-0.96). The range of phenotypic variation coefficient was 3.04%(branch angle)-23.15%(crown width). The correlation coefficient ranged from -0.367(tracheid width and tracheid length-width ratio) to 0.994(diameter of breast height and volume). Principal component analysis showed that the cumulative contribution rate of the four principal components reached 78.46%, which included most of the information about the growth traits and wood traits of the pedigree. The pedigrees were evaluated by growth and wood traits, respectively. Five elite pedigrees with good growth(S78, S81, S80, S84 and S83), five elite pedigrees with good wood quality(S37, S51, S6, S30 and S19), and five elite pedigrees combined with growth and wood traits(S89, S74, S76, S82 and S83) were selected respectively. The selected materials could provide the basis for breeding superior varieties and improving seed orchards construction.

    Identification of Broussonetia papyrifera Transcription Factor BpbZIP1 and Analysis of Its Response to Cadmium Stress
    Sisi CHEN, Muhong XIE, Maokai CUI, Wenkai LI, Zhenggang XU, Caixia JIA, Guiyan YANG
    2022, 42(3):  394-402.  doi:10.7525/j.issn.1673-5102.2022.03.009
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    Heavy metal pollution not only affected the effective area of the soil, restricted the distribution of vegetation, but also harmed the food chain and human health, and cadmium(Cd) pollution was particularly prominent. It was urgent to select plants with strong heavy metal tolerance for bioremediation in tailings areas. For Broussonetia papyrifera was a pioneer tree species in heavy metals polluted soil, In order to explore the molecular mechanism of B. papyrifera in response to heavy metal stress, a basic leucine zipper(bZIP) transcription factor(named BpbZIP1) from B. papyrifera was cloned, and its basic biological information, Cd stress response and the function of transformed yeast were analyzed to predict the role of BpbZIP1 in response to Cd. The results showed that the open reading frame of BpbZIP1 was 1 713 bp, the encoded protein contained 570 amino acids, the molecular weight was 62 902.38 Da, and the theoretical isoelectric point was 4.62. It had a close evolutionary relationship with Arabidopsis thaliana AtbZIP1. Under 150 μmol·L-1 CdCl2 treatment, the BpbZIP1 in B. papyrifera could be induced differently, and the expression of BpbZIP1 in roots was 17.4-fold higher than that in controls at 3 h after induction. Transformation of BpbZIP1 into yeast could significantly improve Cd resistance of transgenic yeasts. When the CdCl2 concentration was higher than 0.6%, the growth vigor of transgenic yeast was 1.54-1.71-fold higher than that in controls. These results suggested that BpbZIP1 gene could positively respond to Cd stress, and its expression benefited Cd stress tolerance. BpbZIP1 was an important candidate in response to Cd stress of B. papyrifera.

    Preliminary Study on Cross Fertility in ‘Kushui’ Rose
    Ying WANG, Yujie ZHANG, Zihui JIAO, Xueli LI, Kai WANG, Xiaowei DA, Kun SUN, Hui ZHANG
    2022, 42(3):  403-411.  doi:10.7525/j.issn.1673-5102.2022.03.010
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    In order to explore the reason why there is no fruit in autumn despite of thousands of flowers of ‘Kushui’ rose(Rosa sertate × R. rugosa Yu. et. Ku.) blossoming in garden, we conducted a genetic research on its cross fertility in this study. The results showed that the selfing of ‘Kushui’ rose did not bear fruit in nature or by artificial pollination. Meanwhile, there were no fruit obtained once pollen of ‘Kushui’ rose being pollinated on the stigmas of the other varieties in Rosa. In contrast to this, ‘Kushui’ rose could produce amount of seeds when pollinated with pollen from the same other materials. All evidences supported that the pollen of ‘Kushui’ rose rose was sterile. Further, several reproductive features, such as stamen petallization, P/O value, pollen heteromorphorsis, and pollen germination rate were studied, with the results showing the pollen of ‘Kushui’ rose deformed so seriously that the germination rate was nearly zero. Furthermore, microscopic observation of pollen mother cells at different phases of meiosis revealed there were so much abnormal behaviors of chromosome allocation as well as unequal cytokinesis that plenty of triads were yielded that coexisting with deformed tetrad in pollen sac. In conclusion, the study shows that the abnormal division of pollen mother cells of ‘Kushui’ rose is responsible for the pollen sterility and no fruit setting when used as paternal parent in hybridization. And that pollen abortion causing male sterile, rather than self-incompatibility commonly distributing in Rosaceae, is responsible for the intraspecific cross infertility of ‘Kushui’ rose, further implying it is feasible to improve its germplasm by acting as a maternal in cross-breeding.

    Molecular biology
    Genome-wide Identification and Expression Analysis of nsLTP Gene Family in Populus trichocarpa
    He CHENG, Shuanghui TIAN, Yang ZHANG, Cong LIU, De’an XIA, Zhigang WEI
    2022, 42(3):  412-423.  doi:10.7525/j.issn.1673-5102.2022.03.011
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    Non-specific lipid transfer proteins(nsLTP), a polygene family of basic lipid transfer proteins, are responsible for the transfer of phospholipids between fatty acids and membrane in vitro, and play an important role in plant growth and development and the response to stress. There have been no reports on the nsLTP family of Populus trichocarpa. In this study, the number of genes, genetic relationship, gene structure and conserved motif of coding proteins of PtrnSLTP family members were analyzed at the genome-wide level. The results showed that the PtrnsLTP family was composed of 39 genes and evolved into five subfamilies, among which six genes in subfamily A, two genes in subfamily B, 13 genes in subfamily C, three genes in subfamily D, and 15 genes in subfamily E. PtrnsLTP family contained seven pairs of paragenetic homologous genes, among which one pair was greater than one, and the six pairs of Ka/Ks were all far less than one. Moreover, these six pairs of genes were all on the same large evolutionary branch, and different evolutionary pressures led to functional differentiation among genes. Both encoded proteins contained conserved motifs in Motif 1 and Motif 2. Using qRT-PCR technology combined with P. trichocarpa data to study the tissue expression of PtrnsLTP and the response characteristics of salt stress, it was found that all family members expressed in the roots, stems and leaves of P. trichocarpa. The results were basically consistent with the prediction results by the bioinformatics analysis of the website. There were 11, 15 and 13 members respectively having higher expression in roots, stems and leaves, indicated that this gene family was involved in the growth and development of different tissues of P. trichocarpa. Under NaCl stress, among the 39 genes in the family, 26 genes expressed in the root and 14 genes expressed in the leaves increased with the increase of stress time, while 32 genes expressed in the stem first increased and then decreased. The results have a positive role in promoting the identification of the biological function of PtrnsLTP family genes and the work of salt stress response gene resources.

    Transcriptome Analysis of Hemerocallis fulva Leaves Respond to Low Temperature Stress
    Dongmei HUANG, Ying CHEN, Lu BAI, Di’an NI, Yiyang XU, Zhiguo ZHANG, Qiaoping QIN
    2022, 42(3):  424-436.  doi:10.7525/j.issn.1673-5102.2022.03.012
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    Low temperature is a kind of abiotic stress in the growth process of Hemerocallis fulva. The transcriptome data of Hemerocallis fulva leaves under cold treatment groups (10, 5, 0 ℃) and control group (15℃) was compared, and a total of 2 457 DEGs were screened, including 1 253 up-regulated genes and 1 204 down-regulated genes. The results showed that differentially expressed genes(DEGs) were mainly concentrated in 49 GO processes such as cellular process, metabolic process and catalytic activity, and 42 KEGG metabolic pathways, including metabolic pathways, biosynthesis of secondary metabolites and plant hormone signal transduction. Among them, the DEGs involved in plant hormone signal transduction pathway changed in varying degrees, GH3.10 gene was increased to 13.624 times higher than the control group, IAA1 gene was down-regulated to 0.120 times; the DEGs involved in soluble sugar synthesis pathway also changed by 0.076-28.114 times. Then, 29 DEGs shared by treatment groups were analyzed by heat map and gene regulation networks. Based on the position of genes in gene regulation networks, the roles of ABCF5OFPs and SWEETs in response to cold were analyzed respectively. The results of this study provide a theoretical basis for further exploring the key genes of low temperature response, germplasm development and molecular breeding of Hemerocallis fulva.

    Chloroplast Genome Structure and Interspecies Relationship of Sanicula rubriflora
    Chen YANG, Xueying YAO, Zhixiang CHEN, Qizhi WANG
    2022, 42(3):  437-445.  doi:10.7525/j.issn.1673-5102.2022.03.013
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    As a kind of medicinal herb, Sanicula rubriflora F. Schmidt is easily confused with other medicinal plants of the same genus after drying and interspecific relationship is controversial. The structure and characteristics of the chloroplast genome of S. rubriflora were reported for the first time, the sequencing data was spliced and annotated using bioinformatics methods, and the chloroplast genome structure and phylogeny were further analyzed. The length of the chloroplast genome sequence of S. rubriflora was 155 721 bp, including a large single copy region(LSC) of 85 981 bp and a small single copy region(SSC) of 17 060 bp, and were separated by inverted repeat sequence(IRs) of 26 340 bp. The GC content of the chloroplast genome of S. rubriflora was 38.20% and contained 129 genes, 84 protein-coding genes, 37 tRNA genes and 8 rRNA genes respectively. The chloroplast genome structure of S. rubriflora was highly conservative. The coding genes contained a total of 51 907 codons, with 5 095 leucine and 689 tryptophan. Simple repeated sequence analysis revealed a total of 32 sites, and most of them were single base repeat A/T type. The clustering results of the chloroplast genome supported that the subfamily Hydrocotyloideae was a relatively primitive group in the Umbelliferae family. The subfamily Saniculoideae and Apioideae were sister groups, which were the more close relationship of the Umbelliferae. Sanicula was a relatively natural group. S. rubriflora and S. flavovirens were related sister species, but their morphology and geographical distribution were quite different. The results provid a basis for the interspecies identification of the genus Sanicula and lay the foundation for further study of its phylogenetic relationship and interspecies evolution.

    Structure of Chloroplast Genome and its Characteristics of Sphaerophysa salsula
    Dandan SU, Yuping LIU, Tao LIU, Changyuan ZHENG, Yu ZHANG, Yanan WANG, Na QIN, Xu SU
    2022, 42(3):  446-454.  doi:10.7525/j.issn.1673-5102.2022.03.014
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    The whole chloroplast genome of Sphaerophysa salsula was sequenced and assembled based on Illumina high-throughput sequencing platform. The assembly result indicated that the whole chloroplast genome of S. salsula had a total length of 123 327 bp, with a loss of IR region. The chloroplast genome encoded a total of 108 genes, including 74 protein-coding genes, 4 rRNA genes and 30 tRNA genes. and 99 SSR loci were found, and among them the number of mononucleotide, dinucleotide and trinucleotide repeat motifs was 75, 17 and 7, respectively. The phylogenetic analysis results showed that S. salsula and Alhagi sparsifolia were sister species, which had the closest genetic relationship. The results would lay the foundation for the genetic diversity, population genetic structure and speciation mechanism of S. salsula in the future.

    Physiology and Ecology
    Effects of Spraying NaHS at Different Growth Stages on H2S Production and Reactive Oxygen Species Metabolism of Naked Oat Leaves under Saline-Alkali Stress
    Jianxin LIU, Ruirui LIU, Xiuli LIU, Haiyan JIA, Ting BU, Na LI
    2022, 42(3):  455-465.  doi:10.7525/j.issn.1673-5102.2022.03.015
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    In order to illuminate the regulating mechanism of gas signal hydrogen sulfide(H2S) on reactive oxygen species(ROS) metabolism of naked oat(Avena nuda) under saline-alkali stress, and to screen and determine the optimal application period and concentration of H2S, the effect of spraying NaHS(H2S donor; 0, 25, 50, 100, 200 and 400 μmol·L-1) at different periods(seedling stage, jointing stage, heading stage, flowering stage and filling stage) on the H2S content, H2S production related enzyme activityof L-cysteine desulfhydrylase(LCD) and ROS metabolism-related enzyme activities and substance contents in leaves of soil-potted naked oat under 3.0 g·kg-1 salt-alkali mixed stress(NaCl∶Na2SO4∶NaHCO3∶Na2CO3 molar ratio was 12∶8∶9∶1) were carried out respectively. The results showed that spraying period, NaHS concentration and their interaction had significant effects on the content of H2S, superoxide anion(), hydrogen peroxide(H2O2), malondialdehyde(MDA), ascorbic acid(AsA) and glutathione(GSH), and the activities of LCD, superoxide dismutase(SOD), catalase(CAT), peroxidase(POD), ascorbic acid peroxidase(APX) and glutathione reductase(GR) in naked oat leaves under salt-alkali stress respectively. Compared with spraying 0 μmol?L-1 NaHS, spraying a certain concentration of NaHS increased the contents of H2S, AsA,GSH and the activities of LCD, SOD, CAT, POD, APX and GR, and reduced the accumulation of , H2O2 and MDA respectively. However, the optimal spraying period and NaHS concentration of the above indexes were different. Comprehensive analysis of membership function showed that spraying 25-200 μmol?L-1 NaHS at the seedling stage and jointing stage had the highest the comprehensive evaluation value(D). It indicated that spraying 25-200 μmol?L-1 NaHS at the seedling-joint stage improved the ROS scavenging ability, alleviated the oxidative damage of salt-alkali stress on naked oats.

    Effects of Different Planting Years on Rhizosphere Soil Physiochemical Properties and Microbial Community of Zanthoxylum bungeanum
    Libin LIAO, Fusun SHI, Nannan ZHANG, Xiaoxia CHEN, Huanhuan BU, Fuyong SUN
    2022, 42(3):  466-474.  doi:10.7525/j.issn.1673-5102.2022.03.016
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    To reveal the effect of different planting years on rhizosphere microecosystem of Zanthoxylum bungeanum, the changes of microbial community structure, physiochemical properties and enzyme activities were studied, and the phospholipid fatty acid(PLFAs) contents of microorganisms, pH, total organic carbon(TOC), available potassium, polyphenol oxidase activity, et al. were investigated in the rhizosphere soils of 1, 15 and 30-year-old Z. bungeanum trees. The results showed that the soil pH and the activity of polyphenol oxidase decreased with the increase of planting years, and the soil available K and P increased significantly at 15 and 30 a. The contents of total PLFA, fungi and gram-negative bacteria(G-) in soil increased with planting years, and reached the maximum at 30 years, which were 401.95, 88.14 and 118.61 μg·g-1, respectively. The contents of gram-positive bacteria(G+) and actinomycetes PLFA increased first and then decreased with planting years. Bacteria/fungi and G+/G- decreased with the increase of planting years. Compared with 1 year plants, bacteria/fungi and G+/G- decreased by 34.14% and 38.87% at 30 years plants respectively. Soil pH was significantly negatively correlated with total PLFA, G-, bacteria and fungi, and positively correlated with G+/G- respectively. The activity of polyphenol oxidase was significantly negatively correlated with fungi, and positively correlated with bacteria/fungi and G+/G- respectively. Z. bungeanum rhizosphere soil physiochemical properties, enzyme activities and microbial community structure changed significantly with the increase of plant years. The changes of rhizosphere soil pH, available phosphorus, available potassium, polyphenol oxidase activity were the main environmental factors that affected the microbial community and composition, the increase of fungi and G- and the decrease of polyphenol oxidase activity might be the important reasons that induced root rot of Z. bungeanum.

    Variation of Flower Color and Pigment Contents of Mirabilis jalapa During Flowering
    Bing ZHOU, Xiaohong YAN, Yihan LEI, Chao PENG, Lihua CHEN
    2022, 42(3):  475-482.  doi:10.7525/j.issn.1673-5102.2022.03.017
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    Mirabilis jalapa, one of the widely distributed garden flowers in China, is rich in color, but the regulating mechanism of pattern and color of M. jalapa with different flower colors during flowering are still unclear. The color phenotype and pigment contents of the corolla of M. jalapa with different flower colors during flowering were which were tested by chromometer and ultraviolet-visible spectrophotometry respectively. The results showed that the corolla of purple red M. jalapa changed from light green to purple red from bud to bloom, and its values of lightness L* and hue b* decreased, but the values of hue a*, chromaticity C* and chromatic triangle h increased, the contents of chlorophyll decreased gradually, but the contents of carotenoid, anthocyanin and total flavonoid increased gradually respectively. The corolla of yellow M. jalapa changed from light green to yellow, had stable chlorophyll and total flavonoid contents, and higher carotenoid contents, its values of chromaticity C*, hue a* and b* at blooming period were the highest. The corolla of white M. jalapa changed from light green to white, its values of lightness L*, chromaticity C*, hue a* and b* and the contents of the chlorophyll at transitional period were the highest, the contents of anthocyanin and total flavonoids were low, but increased gradually with the flowering process, and the contents of carotenoid were stable. It could be concluded that there were differences in the patterns of color changes during the flowering of M. jalapa, which were closely related with the pigment changes.

    Effects of Flooding Stress on Photosynthetic Characteristics of Yulania stellata and Its Cultivars
    Xiwu DU, Jun QIN, Kang YE, Yonghong HU, Yiwei TAO, Yongzheng PENG, Yanxiang SHEN, Yan LIANG, Li ZENG
    2022, 42(3):  483-491.  doi:10.7525/j.issn.1673-5102.2022.03.018
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    In order to clarify the response rules of photosynthetic and physiological characteristics of Yulania stellata to flooding stress, the double-pot method was used to Y. stellata and its 4 cultivars by flooding. The changes in chlorophyll content, specific leaf area, photosynthetic parameters and chlorophyll fluorescence parameters after different flooded days were measured respectively. The results showed that the changing trends of chlorophyll a, chlorophyll b and total chlorophyll were slightly different under flooding stress, and there were no significant changes in Y. ‘(Mag’s) Piroutte’ and Y.stellata ‘Chrysanthemiflora’, while Y.‘Bety’, Y.stellata and Y.‘Judy’ showed a downward trend respectively. The net photosynthetic rate(Pn) showed a downward trend, and the maximum net photosynthetic rate(Pnmax) and light saturation point(LCP) of the treatment groups were significantly lower than those of the control group respectively. The dark respiration rate(Rd) of Y. stellata ‘Chrysanthemiflora’ and Y. ‘(Mag’s)Piroutte’ of the treatment group was higher than that of the control group. The Rd of Y. ‘Bety’, Y. stellata and Y. ‘Judy’ were significantly reduced after flooding treatment to reduce the consumption of photosynthetic products respectively. The effective photochemical quantum yields(ΦPSⅡ) of Y. ‘(Mag’s)Piroutte’ and Y.‘Judy’ decreased by 8.4% and 24.7% under flooding stress; Y.stellata ‘Chrysanthemiflora’, Y.‘Bety’ and Y.stellata decreased significantly, respectively 76.7%, 85.7% and 64.6%. The maximum photochemical quantum yield(Fv/Fm) of each cultivar of Y.stellata. had a downward trend. The photosynthetic characteristics of Y.stellata andits cultivars were affected by flooding stress, but different cultivars had differences. Except for Y.‘Bety’, the Photosynthesis of the other 4 Y.stellata cultivars performed normally during the 7 d before treatment; Y. ‘(Mag’s)Piroutte’ had strong photosynthetic capacity and waterlogging adaptability. The results provid a theoretical basis for the selection of Y.stellata cultivars suitable for planting in wet and waterlogged areas in southern China.

    The Effect of Exogenous Proline on the Stomata of Nitraria tangutorum Leaves under Natural Drought
    Bin WEI, Yi LI, Shiping SU
    2022, 42(3):  492-501.  doi:10.7525/j.issn.1673-5102.2022.03.019
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    Taking Nitraria tangutorum, a perennial desert plant in the field environment, as material, scanning electron microscope was used to observe the microstructure of N. tangutorum leaf stomata under different exogenous proline concentrations, and the length, width, area and density of the leaf epidermal stomatal apparatus were measured respectively, and the difference of the same index under different proline concentration treatments were compared. The concentrations of exogenous proline spraying were set at 50, 100, 150, 200, 250 mg·L-1 respectively(Proline, abbreviated as Pro or P) under natural drought stress, and the drought tolerance of N. tangutorum were tested respectively. The observation results showed that the stomata guard cells of N. tangutorum leaves were kidney-shaped, and the stomata were randomly distributed in the leaf epidermis. The stomatal apparatus was mostly irregular, and the stomata were oval and leaf surface waxy less. And the leaf cells appeared folds and sinks. Compared with the control, under different concentration treatments and different sampling days, the stomata length, width and area decreased, and the stomata density increased. As the concentration increased and the number of sampling days changed, the overall stomata length showed a downward trend. When the concentration reached at 200 mg·L-1, the decreasing amplitude increased, and the overall width and area showed a trend of first decreased, then increased and then decreased, while the stomata density showed an overall upward trend. Among them, the stomata length, width and area were at a concentration of 50 mg·L-1, the number of sampling days reached the maximum on the first day, and the stomatal density reached the maximum at the concentration of 150 mg·L-1 and the ninth day of the sampling days. In addition, the stomata length differed significantly on the first day, the sixth day, and the ninth day under the treatment of different concentrations of proline, while the stomata width and area were not significantly different. Except for the beginning, the stomata density was different due to the different proline concentration. The difference was significant(P<0.05). The results showed that spraying different concentrations of exogenous proline on N. tangutorum leaves under natural drought stress could reduce the length, width and area of the stomata, and increase the density of stomata, could provide a theoretical basis for N. tangutorum drought resistance.

    Effects of Soil Moisture and Light Intensity on Growth and Photosynthetic Characteristics of Pterospermum kingtungense Seedlings
    Shanshan ZHANG, Chunming YUAN
    2022, 42(3):  502-511.  doi:10.7525/j.issn.1673-5102.2022.03.020
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    One-year-old seedlings of Pterospermum kingtungense were selected as materials, effects of soil moisture and light intensity on growth(aboveground biomass, underground biomass, leaf number and plant height) and photosynthetic characteristics(net photosynthetic rate, transpiration rate, stomatal conductance, intercellular carbon dioxide concentration, stomatal limiting value and instantaneous water use efficiency) of P. kingtungense seedlings were investigated respectively. Two-factor of controlled test was analyzed. Six soil moisture gradients(100%, 90%, 80%, 60%, 40% and 30% field capacity) and four light intensity gradients(100%, 75%, 50% and 25%full light) were designed respectively. The results showed that: ①With the decrease of soil moisture, the growth index and photosynthetic characteristics of P. kingtungense seedlings first increased and then decreased, and generally reached the peak in W2(32.11%) or W3(28.54%) treatment. It performed worst under drought stress(W6). ②With the decrease of light intensity, except stomatic limitations(Ls), the four growth indexes and other five photosynthetic characteristics of P. kingtungense seedlings showed a trend of first increasing and then decreasing, and all of them performed best under the light intensity of L2(75% full light) or L3(50% full light). ③The comprehensive evaluation scores of different soil moisture and different light intensity showed that the comprehensive evaluation score of W2 soil moisture or L2 light intensity was the highest respectively, which proved that the appropriate soil water environment and moderate shading were the most suitable for the growth and photosynthesis of P. kingtungense seedlings. ④The results of binary linear regression analysis demonstrated that soil moisture and light intensity had significant effects on the growth and photosynthesis of P. kingtungense seedlings, respectively. This study elucidates the mechanism of soil water and light on regeneration of P. kingtungense seedlings, and could provide a theoretical basis for seedling breeding and population recovery and reconstruction in the future to a certain extent.

    Technique and Method
    Establishment of Agrobacterium rhizogenes-mediated Genetic Transformation System of Eucalyptus urophylla × E. grandis
    Ping LUO, Haonan ZHANG, Jianmin XU, Bing HU, Xiaoping WANG, Guangyou LI, Chunjie FAN
    2022, 42(3):  512-520.  doi:10.7525/j.issn.1673-5102.2022.03.021
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    Agrobacterium rhizogenes-mediated genetic transformation systemplays an important role in the identification of functional genes. To establish Agrobacterium rhizogenes-mediated genetic transformation system in Eucalyptus, the different strains infected the leaves and stems of Eucalyptus urophylla × E. grandis were used to select the appropriate A. rhizogenes strains and explant types, and the effects of Agrobacterium concentration and infection time on hairy root induction were explored. The results showed that MSU440 was the optimal A. rhizogenes strain and leaves was the preferred explants for hairy root induction. The highest hairy root induction rate reached 81.0% with the average root length of hairy roots was 3.23 cm. The leaves were infected by MSU440 which grown to a bacterial density of 0.3(OD600) for 30 min, then they were co-cultured for 48h and transferred the selection medium supplemented with 20 mg·L-1 Kanamycin. The exogenous genes were stably integrated in the genome of Eucalyptus hair roots which confirmed by PCR analyses and GUS staining, and the transformation efficiency reached 20.2%. In our study, an A. rhizogenes-mediated genetic transformation system of Eucalyptus was preliminarily established, which laid a foundation for gene function identification and further transgenic breeding of Eucalyptus.