Table of Content

20 November 2022, Volume 42 Issue 6

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Systematic and Evolutionary



Diversity of Wild Poisonous Plant Resources in Namjagbarwa Region

Pengying CHEN, Chan YANG, Jian LUO

2022, 42(6):  921-929.  doi:10.7525/j.issn.1673-5102.2022.06.001

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In order to rationally protect and explore the wild poisonous plant resources in the Namjagbarwa region， the distribution of families and genera， the composition of life forms， the toxicity， poisonous parts and toxicological effects of plants were analyzed by field investigation， specimen investigation and related literature books inspection. The results showed that there were 77 families， 167 genera and 242 species of wild poisonous plants in the Namjagbarwa region， among which the dominant families mainly included Araceae， Ranunculaceae， Ericaceae， Fabaceae and Asteraceae respectively. The most poisonous plants with herbaceous life form were 138 species in total. Most of the plant toxic parts were poisonous whole grass， a total of 109 species. There were 184 species with mild poisonous plants， including 50 minor toxic plants， there were only five kinds of highly poisonous and three kinds of strong poisonous. There were 19 poisonous nectar source plants， Scrophulariaceae and Ranunculaceae were dominant； in terms of toxicology， most of them were nervous system poisoning. There were many kinds of poisonous plant resources in the Namjagbarwa region， but there was less use of these resources. It was necessary to strengthen the understanding and protection of poisonous plants， and carry out rational development and explroation.



Morphological Characteristics of Spores, Elaters and Oil Bodies from 11 Species of Lophoziaceae in Xinjiang

Aizezi MEIHERIBANHAN, Sulayman MAMTIMIN

2022, 42(6):  930-938.  doi:10.7525/j.issn.1673-5102.2022.06.002

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To improve the characteristics of the spore，elater and oil body of Lophoziaceae and to provide detailed information for classification，the morphology of 11 species of the Lophoziaceae was observed by scanning electron microscopy， 6 species provided for the morphological characteristics of spore and elater，and 9 species provided for oil body. The results showed that：①The spores of 6 species of Lophoziaceae were spherical， near spherical or elliptical with brown， reddish-brown or yellowish brown respectively. The spore size of them were from 10-16 μm belonging to MI type，and their faces near pole were usually concave. The ornamentation of the above spores were verrucose，clavate，spinate，granular or short baculite respectively. ②The elaters of 6 species of Lopholoziaceae were densely bispiral with obtuse both ends，and their cuticle were smooth or granulate.③The oil bodies of 9 species of Lophoziaceae were spherical，elliptical or oval，with the diameter between 3-10 μm. The detailed description and illustration on morphological evidence of spores，elaters and oil bodies of 11 species of Lophoziaceae might provide the elemental data for the systematic taxonomy and evolution of Lophoziaceae.



The Distribution Location and Morphological Characteristics of Special Tracheids in Tetracentron sinense

Xiran LI, Lu YONG, Biao PAN, Xinxin ZHENG, Jiangtao SHI, Ting LI

2022, 42(6):  939-945.  doi:10.7525/j.issn.1673-5102.2022.06.003

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To identify the cell types and analyze the transport mechanism of vascular tissue and explore the cell evolution during tree evolution，a comprehensive anatomical study was carried out on a kind of special tracheids of Tetracentron sinense. After slicing and dissociating， the distribution position and morphological characteristics of the special tracheids of the 34-year-old T. sinense were observed by using optical microscope and scanning electron microscope. The results showed that the special tracheids were arranged from the inside to the outside radially in the horizontal of the tree， and ran through the growth ring boundary. Most of them were in one row， and few were in two rows. And each special tracheid was connected with wood ray cells on one or both sides. In the longitudinal direction， the special tracheids were connected individually or with several cells terminally. There were three main types of special tracheids： fusiform without end wall， fusiform with one sloping end wall or with two sloping end walls. The average length of special tracheids was 286.44 μm. Its cross section was quadrilateral. The average tangential width of special tracheids were 55.22 μm respectively. The average wall thickness of special tracheids was 1.53 μm. The two ends of the special tracheids were closed without perforation. The number of pits on the side wall of special tracheids was large and the pit membrane was clearly visible. There were a large number of bordered pits on the wall in tangential section， which were arranged in opposite， alternated and scalariform pitting. The single pits on the radial wall were rounded or ellipse. There were 2-10 pits， arranged in 1-4 rows in per region. There were no pits between the special tracheids and the normal tracheids on the radial wall. The distribution of special tracheids has certain regularity. Compared with normal tracheids， the special tracheids were much shorter in length， and the tangential width were slightly larger. The type and arrangement of the pits were similar to those of the pits on the wall of general hardwood species. The shape of the special tracheid was different from the general vessel in the hardwood， and it was in an intermediate state of the evolution from tracheid to vessel， indicated a certain level of evolution of this type of cell.



Sporogenesis and Gametogenesis of Akebia trifoliata

Jie BAI, Tao ZHOU, Jiang MA, Wen LIU, Zhiguo JIANG, Faju CHEN

2022, 42(6):  946-955.  doi:10.7525/j.issn.1673-5102.2022.06.004

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Akebia trifoliata belongs to Akebia of the Lardizabalaceae family， and it is a dioecious， monoecious woody vine， with economic value for medicine and food. In order to understand the sexual reproduction process of A. trifoliate， analyze the reproductive reasons of the low seed setting rate of this species under natural conditions， and provide a theoretical basis for its hybrid breeding and new variety cultivation，the embryological characteristics of mega-and microsporogenesis and female and male gametophyte development of A. trifoliate were observed by paraffin section technology. The main results showed that A. trifoliata had six stamens with four locules in each， and the anther wall was fully differentiated， and it was composed of epidermis（1 layer）， endothecium（1 layer）， middle layers（2-3 layer） and tapetum（1 layer） from outside to inside， tapetum belonged to glandular type respectively. The cytokinesis of the microspore mother cell in meiosis was of simultaneous type， and the tetrads were arranged in a tetredraltah， mature pollen grain was two-celled. There were little pollen abortion， and abortion occurred at the microspore tetrad stage. The ovary was one locular， parietal placentation， ovules were hemianatropous， bitegminous and crassinucellate. Its megaspore was of monosporogenesis type， and tetrads were linearly arranged. The chalazal end megaspore developed into functional megaspores， which underwent three times consecutive mitosis and further developed into polygonum embryo sac with seven cells and eight nucleus，another three megaspores degenerated. The early development of the stamens in the female flowers was normal， but the microspores did not develop further until the uninucleate pollen grain stage， the tapetum was not completely degraded， and the anther wall did not crack， the degeneration of the carpel of the male flowers was due to the early lack of development，the carpels heal incompletely， and no ovules are produced.The process of the mega-microsporogenesis and development of female and male gametophyte of A. trifoliata were normal， which was not the reason for the low seed setting rate， but may be related to external factors affecting pollination.



Genetic Structure and Phylogeography Analysis of Artemisia intramongolica Based on Single Copy Nuclear Genes

Siru TANG, Tiejuan WANG, Fang XIU, Mengyao LIU, Hui ZHANG

2022, 42(6):  956-963.  doi:10.7525/j.issn.1673-5102.2022.06.005

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To investigate the genetic and phylogeography structure of Artemisia intramongolica， which was an important sand-fixing plant in Hunsandak sandy land in China， the homologous single-copy nuclear genes c9065 and c7847 were developed by transcriptome sequencing， and then eight natural populations of A. intramongolica were studied. The results showed that the total length of c9065 and c7847 were 485 bp and 457 bp， respectively， with 14 and 19 variable loci， and 48 and 40 haplotypes were obtained. Based on c9065 and c7847， the haplotype diversity（H_d） was 0.871 6 and 0.934 8， respectively， and all populations were above 0.8. The total nucleotide diversity（π） was 0.008 2 and 0.005 9， respectively， and all populations were above 0.005， indicating that both species and populations had high genetic diversity. The results of AMOVA analysis showed that 99.398% and 98.908% of genetic variation existed within the populations of A. intramongolica based on c9065 and c7847， respectively， with gene flow（N_m）of 6.810 and 7.270， respectively， much greater than 1， indicating that there was extensive gene exchange among populations of A. intramongolica. Based on c9065， N_st<G_st， although N_st>G_st by c7847， but the difference was not significant（P>0.05）， indicating that A.intramongolica population did not exist phylogeographic structure. The neutral test showed that Tajima’s D values for both nuclear geneswere all not significant（P>0.05）， while Fu’s F_s values were all significant（P<0.05）， and combined with the mismatch distribution analysis， A.intramongolica experienced limited population expansion under the neutral mode.



Comparison of Codon Characteristics and Phylogeny of Chloroplast Genome between Ranunculaceae and Paeoniaceae

Yingtong MU, Dongchang FAN, Lijuan LÜ, Xiajie LI, Jingshi LU, Xiaoming ZHANG

2022, 42(6):  964-975.  doi:10.7525/j.issn.1673-5102.2022.06.006

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To clarify the codon usage pattern and phylogenetic relationship of the chloroplast genome between Ranunculaceae and Paeoniaceae， 33 species of Ranunculaceae and seven species of Paeoniaceae were used as materials， and the analysis software CodonW， online software CUSP， and R software were used to analyze the codon characteristics of chloroplast genes， while the MAFFT software， MEGA software were used for phylogenetic analysis respectively. The results showed that the chloroplast genomes of Paeoniaceae and Ranunculaceae （except Columbine） had a high consistency of high frequency codons， with 29 high frequency codons， which were basically biased towards ending with A/U， but the optimal codon were different. The codon bias of the chloroplast genome of Ranunculaceae and Paeoniaceae was mainly influenced by natural selection， and the codon bias of the chloroplast genome of Paeoniaceae was more affected by natural selection than that of Ranunculaceae. The phylogenetic relationship based on the complete chloroplast genome sequence and the CDS sequence of the chloroplast genome showed that although the phylogenetic relationship based on the complete chloroplast genome sequence and the chloroplast genome CDS sequence of Paeoniaceae was different， they could be divided into the peony group and the peony group. The phylogenetic relationship of Ranunculaceae based on the entire chloroplast genome sequence did not conform to the classification relationship of Chinese flora， but it supported the division of Ranunculaceae into four subfamilies and 14 families. The analysis results supported that Paeoniaceae were independent from Ranunculaceae and Ranunculus， and were classified into Saxifraga.

Molecular biology



Study on Adaptive Differentiation of Transcription Factor bHLH94 Gene to Altitude in Hippophae neurocarpa and H.tibetana

Ting QIAN, Fan ZHAO, Yujie ZHANG, Xueli LI, Kun SUN, Hui ZHANG

2022, 42(6):  976-985.  doi:10.7525/j.issn.1673-5102.2022.06.007

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The bHLH proteins are the second large family of transcription factors， which play important roles in plant growth development， physiological metabolism and stress responses. In this study， Hippophae neurocarpa and H.tibetana were used as materials， the positive selection of the transcription factor bHLH94 genewas screened out using transcriptome sequencing. Based on the analysis of gene sequence and expression of HnbHLH94 and HtbHLH94， the response mechanism of bHLH94 genein H.neurocarpa and H.tibetana adapted to altitude was studied. The HnbHLH94 and HtbHLH94 gene encoded 338 and 335 amino acids respectively. Sanger sequencing method verified the correctness of sequences， however there were 10 non-synonymous mutation sites outside the DNA binding domain of the two genes， which was speculated to be related to the adaptive evolution of the gene. As the altitude increased， the HnbHLH94 gene expression decreased， while the HtbHLH94 gene expression increased based on qRT-PCR technology. It suggested that gene might provide a molecular basis for adaptation to altitude， drought， freezing and radiation respectively， and the sequence structure and expression of HnbHLH94 and HtbHLH94 genes responded to the habitat conditions of elevation.



Cloning and Functional Analysis of miR398a from Chrysanthemum× grandiflora in Response to Salt Stress

Liran YUE, Yingjie LIU, Chenxu LIU, Yunwei ZHOU

2022, 42(6):  986-996.  doi:10.7525/j.issn.1673-5102.2022.06.008

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miRNAs played an important role in abiotic stress， and miR398a and pre-miR398a sequences were obtained from Small RNA database of Chrysanthemum×grandiflora. The sequence alignment showed that the cgr-miR398a was highly conserved with the identified miR398a sequences of other plants. Target genes prediction of cgr-miR398a showed that their annotation of targets include superoxide dismutase［Cu-Zn］（CSD2）， copper chaperone（CCS）， A20/AN1-like zinc finger family protein（SAP8）. The qPCR assay showed that the expression levels of miR398a was significantly decreased while target genes showed contrary trend during salt stress in Chrysanthemum×grandiflora. To explore the function of cgr-mir398a in response to salt stress， and an overexpression vector of cgr-mir398a was constructed and transformedinto Arabidopsis thaliana.The result showed that overexpression of the cgr-MIR398a in Arabidopsis could decrease seed germination under salt stress and salt tolerance at the adult seedling stage， therefore cgr-miR398a played a negative regulatory role in Arabidopsis in response to salt stress. The results provided a valuable reference for further exploring the functional mechanism of miR398a during salt stress in Chrysanthemum×grandiflora.



Cloning and Expression Analysis of DELLA gene HbRGL1 from Hevea brasiliensis Müll. Arg

Bi QIN, Mingyang LIU, Meng WANG, Lifeng WANG, Fei HUANG

2022, 42(6):  997-1004.  doi:10.7525/j.issn.1673-5102.2022.06.009

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DELLA protein belongs to the plant-specific GRAS family and is a negative regulator of plant growth. To reveal the molecular regulation mechanism of DELLA gene in rubber tree growth and development， the full-length cDNA sequence of HbRGL1 was cloned from rubber tree cultivar Reyan 7-33-97. The ORF of HbRGL1 was 1 851 bp， and encoded 616 amino acids. Bioinformatics analysis showed that HbRGL1 was an unstable hydrophilic protein containing two conserved domains of DELLA and GRAS， and localized in the nucleus. Phylogenetic analysis showed that HbRGL1 was clustered with HbGAIL of rubber tree， MeGAIL of Manihot esculenta， RcGAI of Ricinus communis， and JcGAIof Jatropha curcas. qRT-PCR analysis showed that the expression level of HbRGL1 in different tissues of rubber treewas significantly different， and its expression level was the highest in flower. HbRGL1 expression was significantly up-regulated in leaves treated by different hormones such as auxin（IAA）， ethephon（ET） and abscisic acid（ABA）. Especially， and it rapidly responded to GA₃ treatments， and had the highest up-regulation of expression level. The study provided theoretical basis for further elucidating the function of HbRGL1 in the growth and development of rubber tree.



Cloning and Drought Tolerance Expression Analysis of PtrWRKY51 Gene in Populus trichocarpa

Anying HUANG, Dean XIA, Yang ZHANG, Dongchen NA, Qing YAN, Zhigang WEI

2022, 42(6):  1005-1013.  doi:10.7525/j.issn.1673-5102.2022.06.010

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In order to clarify the function of PtrWRKY51 gene of Populus trichocarpa， the CDS of PtrWRKY51 gene was cloned using P. trichocarpa from Nisqually-1 strain as material. The function of PtrWRKY51 gene was studied by bioinformatics analysis， and assayed by yeast self-activation verification， and subcellular localization and simulated drought stress by real-time qPCR. The results showed that the CDS of PtrWRKY51 was 579 bp and encoded 192 aa. Bioinformatics analysis and subcellular localization experiments showed that PtrWRKY51 was a non-transmembrane alkaline unstable hydrophilic protein， located in the nucleus and contained a conserved domain unique to WRKY family， it was a WRKY transcription factor classified to class IIc. The yeast self-activation verification experiment showed that PtrWRKY51 gene could self-activation activity. qRT-PCR showed that the expression of this gene was significantly induced by 8% PEG6000 and reached the highest expression in stem and leaf after 12 h stress， while the roots appeared after 24 h stress. This study could provide reference for further study on stress tolerance and biological function of PtrWRKY51.



Sequencing and Analysis of miRNAs in Vegetative and Reproductive Growths of Ceratopteris pteridoides

Jun SUN, Guisheng LI

2022, 42(6):  1014-1022.  doi:10.7525/j.issn.1673-5102.2022.06.011

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Stress Tolerance Functional Analysis of the High Expression Heat Shock Protein HbHSP90.4 Gene from the Latex of Hevea brasiliensis

Huafeng CHEN, Longjun DAI, Mingyang LIU, Bingbing GUO, Hong YANG, Lifeng WANG

2022, 42(6):  1023-1032.  doi:10.7525/j.issn.1673-5102.2022.06.012

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In order to analyze the role of Heat Shock Protein 90（HSP90） in stress and phytohormone signaling transduction in rubber tree， the full-length cDNA sequence of the HbHSP90.4 gene was cloned from rubber tree Reyan73397 latex by PCR， and HbHSP90.4 gene contained a 2 451 bp open reading frame（ORF） and encoded 816 amino acids. The results of bioinformatics analysis showed that HbHSP90.4 contained the HSP90 superfamily and HATPase superfamily domains， which was a member of the HSP90 family. Phylogenetic analysis revealed that this protein was closely related to cassava MeHSP90. Subcellular localization predictions showed that the HbHSP90.4 gene was localized in endoplasmic reticulum. The qRT-PCR results showed that the HbHSP90.4 gene was mainly expressed in rubber latex. Drought， cold， powdery mildew infection， H₂O₂ and MeJA treatments promoted the up-regulation of the HbHSP90.4 gene in latex respectively， while its expression was significantly down-regulated in ETH， SA and ABA treatments respectively. The plant expression vector HbHSP90.4-mScarlet was also constructed to prepare the material for further transgenic plants. This study laid a solid foundation for the elucidation of the molecular regulation of the latex HbHSP90.4 gene in response to rubber tree stress and the phytohormone signaling pathway.



Identification and Analysis of the Potato StCRKs Gene Family and Expression Patterns in Response to Stress Signals

Denggao LI, Rui LIN, Qinghui MU, Na ZHOU, Yanru ZHANG, Wei BAI

2022, 42(6):  1033-1043.  doi:10.7525/j.issn.1673-5102.2022.06.013

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A total of 18 new StCRKs（cysteine-rich receptor-like kinases， CRKs） family genes were identified in potato genome using online tools such as Pfam. These StCRKs localize on chromosome 1， 2， 11 and 12， and all contain conserved CRK domains. In the promoter regions of these StCRKs genes， there are five plant hormones responsive elements， the circadian control element， biotic and abiotic responsive elements， and seed-specific regulation element. The tissue-specific expression pattern were measured using qRT-PCR method in root， stem， leave， and flower tissues in potato during flowering time. The results showed different StCRKs expressed in different tissues. After salicylic acid analogue BTH and 4 ℃ treatment， it was found that 10 StCRKs genes could be induced by BTH and 13 StCRKs genes were able to respond to the low temperature signal. The results provide clues for further elucidation functions of StCRKs on biotic and abiotic stresses.



Cloning and Osmotic Stress Response Analysis of ThGRF2 from Tamarix hispida

Jing WU, Yuanyuan WANG, Danni WANG, Baichao LIU, Zhongyuan LIU

2022, 42(6):  1044-1051.  doi:10.7525/j.issn.1673-5102.2022.06.014

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14-3-3 protein，also called general regulatory factors（GRF），is a type of serine and threonine phosphorylation binding protein， which participates in the regulation of series of physiological processes， such as intracellular basis， signal transduction， plant growth and development， environmental stress response， by interacting with other transcription factors or signaling proteins. In this study， the ThGRF2 gene that was differentially expressed under drought stress and cloned from the drought transcriptome of Tamarix hispida. The CDS fragment of ThGRF2 gene was 786 bp， encoded 261 amino acids. The relative molecular weight was 29.40 kDa， and the theoretical isoelectric point was 4.76. The overexpression vector pROKII-ThGRF2 was constructed， and transformed into T. hispida. The physiological index results showed that ThGRF2 overexpression increased the chlorophyll content， SOD and POD activities， and reduced malondialdehyde content（MDA）， electrical conductivity（EL） and water loss rate under osmotic stress， indicated that ThGRF2 gene played an important role in the response to osmotic stress of T.hispida. It laid a foundation for further exploration of the abiotic stress tolerance function of ThGRF2 gene.

Physiology and Ecology



Quantitative Analysis of Leaf Photosynthetic Heterogeneity Based on Chlorophyll Fluorescence Imaging and Distribution Characteristics of Fluorescence Parameters

Wenhai HU, Yi’an XIAO

2022, 42(6):  1052-1061.  doi:10.7525/j.issn.1673-5102.2022.06.015

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Leaf photosynthesis has high spatial heterogeneity， and chlorophyll fluorescence imaging technology provides convenience for the study of leaf photosynthetic heterogeneity， however， the analytical method for quantifying photosynthetic heterogeneity has not been widely used yet. In this study， the chlorophyll fluorescence images for the sunny and shady leaves of Ficus microcarpa in the mid subtropical region during overwintering period were obtained by using the PAM chlorophyll fluorescence imaging system（Walz， Germany）， and the photosynthetic heterogeneity characteristics of sunny and shady leaves were quantitatively analyzed respectively. The results showed that the degree of photosynthetic heterogeneity and photoinhibition of sunny leaves were significantly higher than those of shady leaves for F. microcarpa during overwintering， and the coefficient of variation（C_V） could be used as a quantitative index of photosynthetic heterogeneity. For sunny leaves 4.30% necrotic areas， 53.30% severe photoinhibition areas（0<F_v/F_m<0.627） and 42.27% mild photoinhibition leaf areas（0.627≤F_v/F_m<0.800） were observed under low temperature but high light condition， respectively. On the other hand， for shady leaves lesser photoinhibition and leaf necrosis were observed under low temperature and low light condition. Furthermore， the quantitative analysis of other fluorescence parameters， such as effective quantum yield of photosynthetic system Ⅱ（PSⅡ） photochemistry（Y（Ⅱ））， quantum yield of regulated energy dissipation（Y（NPQ）） and quantum yield of non-regulated energy dissipation （Y（NO））， showed that the sunny leaves possessed relatively higher photochemical ability， while the shady leaves had relatively higher heat dissipation capacity. In addition， although in winter the high light would lead to the serious accumulation of excitation pressure of the PSⅡ in sunny leaves， which indicated potential risk of serious photoinhibition stress， the necrotic areas were not large， and there were still some low-risk areas with low excitation pressure in sunny leaves. However， the shady leaves under low temperature and low light were mainly low-risk areas with low excitation pressure.



Effects of Selenium Treatment on Physiology and Primary Metabolism of Astragalus adsurgens

Wanting JIANG, Nan YANG, Chen CHEN, Xiaorui GUO, Zhonghua TANG, Hongzheng WANG

2022, 42(6):  1062-1069.  doi:10.7525/j.issn.1673-5102.2022.06.016

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In order to clarify the effects of different concentration Se treatments on the growth， development， photosynthesis and the primary metabolism of plant， Astragalus adsurgens， with high Se accumulation capacity under hydroponic condition was used as material. The results showed that different concentrations of Se had different effects on the growth and development of A. adsurgens. Low concentration Se treatment（5 μmol·L^-1） significantly promoted the growth of A. adsurgens， whereas high concentration Se treatment（100 μmol·L^-1） had the opposite effect. The content of Se in roots and shoots was positively correlated with the treatment concentration， and the transport coefficient of Se in A. adsurgens decreased gradually with the increase of selenium concentration. The results of photosynthetic pigment content and leaf chlorophyll fluorescence parameters showed that low-concentrations of Se promoted photosynthesis of A. adsurgens， while high-concentrations of Se significantly inhibited it， the Se transport coefficient under 100 μmol·L^-1 treatment decreased by 83.5% compared with 5 μmol·L^-1 treatment. After detection and analysis of primary metabolites， it was found that low concentrations of Se up-regulated the metabolic pathways related to amino acid metabolism in plant roots and shoots， while high concentrations of Se mainly up-regulated the metabolic levels of primary metabolites that were closely related to the synthesis of secondary metabolites. Our results showed that treatment with low concentration of Se increased the photosynthesis level of A. adsurgens and the level of primary metabolism related to growth， and promoted plant growth； treatment with high concentration Se increased the level of A. adsurgens by reducing the transport of Se to the ground and more primary metabolites for the synthesis of secondary metabolites were used， and the tolerance of plants to Se stress was improved.



Differences in Leaf Metabolism of Wild Rosa acicularis and Rosa acicularis ‘Luhe’ Based on GC-MS

Yan WU, Sai LI, Kexin WU, Liqiang MU

2022, 42(6):  1070-1078.  doi:10.7525/j.issn.1673-5102.2022.06.017

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In order to explore the differences in the metabolites and metabolic pathways of leaves from wild Rosa acicularis and R. acicularis‘Luhe’，a GC-MS non-targeted metabolomics test was carried out on the leaves of two kinds of R. acicularis，6 amino acids，12 sugars，four alkanes，17 organic acids，five esters and seven other compounds and a total of 51 metabolites were detected respectively. The PCA model was in line with expectations and the OPLS-DA model screened out 19 significantly different metabolites. The analysis of differential metabolic pathways revealed a total of five major differential metabolic pathways. Based on the results， there were significant differences in the metabolism of some organic acids and sugars in as well as in the ABC transporter， phenylpropane biosynthesis， benzoic acid degradation and the metabolism of alanine， aspartic acid and glutamic acid in the leaves of the two species of R. acicularis. It was speculated that there were significant differences in leaf photosynthetic efficiency and resistance to stress， but further study was needed in combination with genome， proteome， transcriptome and other methods.



Responses of Supplemental UV-B Radiation to Physiological Properties and Secondary Metabolites of Thlaspi arvense

Zhihui HUANG, Yining ZHANG, Nana LI, Baojiang ZHENG, Yuhong ZHANG

2022, 42(6):  1079-1087.  doi:10.7525/j.issn.1673-5102.2022.06.018

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In order to understand the growth rule of the medicinal plant Thlaspi arvense under the enhanced ultraviolet（UV-B） radiation， the 2-months-old seedlings of T. arvense were taken as materials， two different radiation intensities were supplemented artificially with 3.26 μW·cm^-2（T1） and 9.78 μW·cm^-2（T2） on the basis of natural light， and the T. arvense grown under natural light as the control， to investigate the response of various physiological indicators and secondary metabolites to different UV-B radiation intensities， such as photosynthetic indicators and chlorophyll fluorescence parameters， photosynthetic pigments（chlorophyll a and b） content， osmotic adjustment substances（malondialdehyde， soluble protein， soluble sugar and proline） content， activity of antioxidant enzymes（SOD， POD， CAT and APX）， and the content of total flavonoids， total phenols and sinigrin. The results showed： net photosynthetic rate， stomatal conductance， transpiration rate， intercellular CO₂ concentration， maximum fluorescence（F_m）， PSⅡ maximum photochemical efficiency（F_v /F_m） and Potential Photochemical Efficiency of PSⅡ at Two Radiation Intensities（F_v /F₀） and other chlorophyll fluorescence parameters decreased with the increasing of radiation intensity respectively. However， the chlorophyll a and chlorophyll b， osmotic adjustment substances， such as soluble protein， soluble sugar and proline， superoxide dismutase（SOD）， peroxidase（POD）， catalase（CAT） and ascorbate catalase（APX）， total phenols and sinigrin firstly increased and then decreased with the increase of radiation time， while the total flavonoids content gradually increased， which was significantly different（P＜0.05） from the control in the late radiation period. According to the comprehensive analysis of the research results， under different radiation intensities， T. arvense might improve the synthesis of total flavonoids and total phenols， the accumulation of soluble protein and soluble sugar， and the content of chlorophyll， and the activity of superoxide dismutase and peroxidase and catalase increased， to resist the influence of ultraviolet stress， and then showed the resistance and the adaptability to the stress environment of adversity. The results provided a theoretical basis for breeding and further utilization of T. arvense.



Effects of Exogenous NO on Soil Nutrient Content and Seedling Growth Characteristics from Potted Catharanthus roseus under Light-shading Stress

Ying LIU, Jiayi WU, Ling JIN, Qianru JI, Yujie FU, Dewen LI

2022, 42(6):  1088-1095.  doi:10.7525/j.issn.1673-5102.2022.06.019

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To explore the effect of exogenous NO donor（sodium nitroprusside， SNP） on soil nutrient content and Catharanthus roseus seedlings growth under light-shading stress， four treatments（normal culture， external application of SNP， light-shading treatment and external application of SNP+ shading treatment） were designed， and the change characteristics of C， N， P nutrient contents and physicochemical index from potted soil， and the seedling shoot growth index（plant height， leaf length， leaf width， leaf area， fresh mass and dry mass） were analyzed respectively. The results showed that soil water content， soil pH and soil organic carbon content increased significantly（P<0.05） under the combination of shading stress and exogenous NO， while the contents of total nitrogen， alkali-hydrolyzable nitrogen， total phosphorus and available phosphorus in soil increased but not significantly， and the soil C/N and C/P values reached significant level， and the C/N value increased significantly（P<0.05）； Correlation analysis showed that there was a positive correlation between soil pH value and organic carbon content（P<0.05）， the change of soil organic carbon content was positively correlated with plant height and leaf fresh mass（P<0.05）. It indicated that external application of SNP could promote the increase of soil pH under light-shading stress， change soil organic carbon content， increase C/N value， improve soil fertility， and promote leaf growth and development， then increase the biomass of C. roseus， and provide basis for scientific balanced fertilization and alkaloid content accumulation of C. roseus.



Effects of Shading on Photosynthetic Characteristics and Non-structural Carbohydrate Content of Heptacodium miconioides Seedlings

Chunyan WEI, Yueling LI, Zexin JIN, Guangyu LUO, Chao CHEN, Fangquan SHAN

2022, 42(6):  1096-1105.  doi:10.7525/j.issn.1673-5102.2022.06.020

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To investigate the growth and adaptation of Heptacodium miconioides under different light conditions， two-year-old H. miconioides seedlings were used as materials under three light intensities（full light， 60% shade and 90% shade）， and the photosynthetic characteristics and non-structural carbohydrate（NSC） content of H. miconioides were measured. The results showed that： （1）The diurnal changes of net photosynthetic rate（P_n） of H. miconioides seedlings under three light intensities showed “single-peak” curves. The daily mean values of P_n under three light intensities ranked as full light>60% shade>90% shade. The daily mean values of water use efficiency（W_UE） under 90% shade treatment was significantly lower than those under full light and 60% shade treatments； （2）Compared with full light， the light compensation point（L_CP） and dark respiration rate（R_d） of H. miconioides seedlings under 60% shade decreased significantly， while the light saturation point （L_SP）， maximum net photosynthetic rate（P_nmax）， maximum carboxylation rate（V_cmax）， maximum electron transport rate（J_max）， and triose phosphate utilization rate（T_PU） had no significant difference respectively. As shading degree increased， the L_SP， P_nmax， L_CP， R_d， V_cmax， J_max and T_PU of H. miconioides seedlings under 90% shade were significantly lower than those under full light and 60% shade treatments. （3）The contents of starch， soluble sugar and NSC in leaves of H. miconioides seedlings were highest under 60% shade treatment， and decreased significantly under 90% shade treatment. The NSC content of H. miconioides seedlings was significantly positively correlated with P_n， V_cmax， J_max and T_PU respectively. Therefore， H. miconioides had ability of photopholic and shade-tolerant at the seedling stage， and plants could adjust the photosynthetic characteristics of leaves by reducing L_CP and R_d， and maintained a high NSC content under moderate shading conditions， thereby to improve the utilization rate of weak light and carbon assimilation ability， however， excessive shading was not conducive to the growth of H. miconioides seedlings.

Plant biochemistry



Histochemical Localization of Arabinogalactan Proteins During Fruit Development from Ziziphus jujuba ‘Lingwu Changzao’

Jing WANG, Yingcai ZHANG, Shanshan TAO

2022, 42(6):  1106-1120.  doi:10.7525/j.issn.1673-5102.2022.06.021

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To reveal the distribution of Arabinogalactan proteins（AGPs）of Ziziphus jujuba ‘Lingwu Changzao’during fruit development， fruits in the early bulking period， rapid enlargement period， coloring period and maturation period were used as materials and the distribution of AGPs were identified by methods of histochemistry and immunofluorescence localization respectively. The results showed the brown red precipitate formed by βGlcY-AGPs and antigen recognized by MAC204 antibody distributed in the cell walls and cell interiors of the epicarp and the alongside internal layers of closely arranged mesocarp small cells in different periods fruit. The brown red precipitate formed by βGlcY-AGPs and antigen recognized by MAC204 antibody distributed in the cell walls and the inner part of the large ovoid parenchyma cells in the inner mesocarp during the early bulking period， while they were mainly distributed in the parenchyma cell walls during the rapid enlargement period， coloring period and maturation periods， with no distribution in the inner part of most cells. With the ripening of fruits， the mesocarp parenchyma cells became more loosely arranged with elongated intercellular spaces， cell rupture occurred， and the distribution of brown red precipitate formed by βGlcY-AGPs and antigen recognized by MAC204 antibody gradually decreased. The brown red precipitate formed by βGlcY-AGPs and antigen recognized by MAC204 antibody distributed in the cell walls and cell interiors of all cells in the vascular bundle sheaths， xylem， phloem， and cambium of vascular bundles in different periods fruit respectively， and the number and size of vascular bundles gradually decreased with fruit development and further increased in size， and the distribution of brown red precipitate formed by βGlcY-AGPs and antigen recognized by MAC204 antibody gradually decreased. The results suggested that βGlcY and MAC204 were respectively effective method for histochemical localization and good antibody for immunofluorescence of Z. jujuba ‘Lingwu Changzao’ fruit， and there were some differences in brown red precipitate formed by βGlcY and the fluorescence intensity of antigen recognized by MAC204 antibody in different tissues of fruits at different periods， AGPs probably participated in morphogenesis during vascular bundles development of Z. jujuba ‘Lingwu Changzao’ fruit， which provided protection and nutritional support and polysaccharide accumulation for fruit development.