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    20 March 2021, Volume 41 Issue 2
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    A New Species and a New Variety of Delphinium(Ranunculaceae) from China
    Wen-Tsai WANG
    2021, 41(2):  161-163.  doi:10.7525/j.issn.1673-5102.2021.02.001
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    A species， belonging to Sect.Delphinastrum DC. subsect. Grandiflora W.T.Wang， and a variety of D. iliense Huth were described as new.

    Oreorchis bashanensis Wang Yong, a New Species of Orchidaceae from Shaanxi, China
    Yong WANG
    2021, 41(2):  164-167.  doi:10.7525/j.issn.1673-5102.2021.02.002
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    A new species of Orchidaceae， Oreorchis bashanensis Wang Yong from Dabashan mountain area in Shaanxi was reported. The species was closely related to O.patens. Comparing with O.patens， the new species was decedious. The leaves fell during its flowering period. During its nutritional growth period， there were 2-3 leaves on each individual. It had larger pseudobulb and leaf blade. The lateral petals were erect. The claw of lip petal was about half the length of the whole lip petal. The orange callus protruded between lateral lobes on lip. However， O.patens was evergreen， and it had 1-2 leaves on each individual. The claw of lip petal was about 1/4 the length of the whole. The lateral petals were curved. The longitudinal lamellae was white or pale yellow. The type specimens were deposited in Herbarium of Shaanxi University of Technology（HZTC）.

    Phenological Observation of the Extremely Small Population Plant Styrax zhejiangensis
    Guo-Lin FU, Ting-Ting LI, Ye-Hua ZHANG, Xiang-Ying WEN, Xue-Chu MA, Chu-Ping WU
    2021, 41(2):  168-173.  doi:10.7525/j.issn.1673-5102.2021.02.003
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    Styrax zhejiangensis was a species with extremely small population， and endemic in Taohuawu， Longjiang， Jiande Forest Farm， Zhejiang Province. To clarify the reason this species endangered， phenology of S.zhejiangensis was observed， including its flowering and seed setting pattern， as well as the biological characteristics of seeds. The results showed that flowers and leaves shared the same proliferation period. The characteristics of flower organs and pollen were similar to those plants of the same genus， and the seed setting rate was 21%-32%. Seeds had high morphological variations， and their insufficient plumpness and poor water permeability caused by dense and hard seed coat may affect seed germination. Abundant rainfall， and temperature differences between day and night， were two major drivers that would aggravate the falling of flowers and fruits. More attention should be paid to their seeds， flowers and fruits if we aim to conserve this endangered species.

    Responses of Water Absorption and Germination of Angelica sinensis Seeds to Winged Perianth Removal
    Yong-Wei MI, Cheng-Wen GONG, Guo-Xiang WANG, Yong-Hong QI
    2021, 41(2):  174-179.  doi:10.7525/j.issn.1673-5102.2021.02.004
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    The water absorption and germination process for intact and de-winged seeds（winged perianth removed） of Angelica sinensis were determined to provide a theoretical basis for its artificial propagation. With intact and de-winged seeds of A. sinensis， and intact seeds being used as a control group（CK）， the 1000-grain weight， water content， volume， volume weight and water absorption ratio of seeds were measured， in the meanwhile， the germination test was proceeded under natural room temperature and in laboratory. Compare with CK， 1000-grain weight and volume of de-winged seeds decreased by 46.87% and 90.78%（P<0.01）， respectively. The volume weight and water content of de-winged seeds increased by 475.92%（P<0.01） and 20.15%（P<0.05）， respectively. The time ofwaterabsorption to reach stationary phase of de-winged seeds was 2.48 h， and was shorter than that of CK by 3.38 h， because the winged perianths significantly inhibited the waterabsorptioncapacity of seeds， and the soaking process fitted the logistic equation. Compared with CK， the germination potential， germination rate and germination index of de-winged seeds increased by 256.70%， 12.28% and 27.77%（P<0.01）， respectively. The volume and weight of A. sinensis seeds were decreased， the water absorption capacity and germination quality of seeds were improve by removing winged perianths. Therefore， the de-winged seeds of A.sinensis should be applied in agricultural production.

    Effects of Micro-habitats on Water Retention and Loss of a Drought-tolerant Saxicolous Moss Grimmia pilifera
    Dong QIU, Jing ZHANG, Nan WU, Ye TAO
    2021, 41(2):  180-190.  doi:10.7525/j.issn.1673-5102.2021.02.005
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    The Grimmia pilifera is a typical drought-tolerant saxicolous moss species， and the water is the key factor influencing its survival and reproduction. To explore the effects of micro-habitats to water physiology of G.pilifera， three typical micro-habitats（habitat S-1 is the bare rock shaded by bamboo individuals； habitat E is the exposed rock； habitat S-2 is the bare rock shaded by shrub Ficus pumila） were chosen in the Dalongshan National Forest Park in Anhui Province， then the differences in individual plant size， saturated water content and water content indices of G.pilifera during dehydration in different micro-habitats， and finally the impact of micro-habitats on moss water stress tolerance and trade-off characteristics were comprehensively assessed. It was found that the plant size and saturated water content indices varied significantly among three micro-habitats； there into， the moss plant size in habitat S-1 was low but the internal and external water contents were relatively high； the plant sizes and external water contents in habitat E and habitat S-2 were similar but the external water content was much more higher of the former. The external water contents of moss plants in the two shaded habitats S-1 and S-2 were seven times the internal water contents， and their ratios were notably higher than that in habitat E. The water content indices of G.pilifera in habitat E the same time during dehydration were higher than that in the two shaded habitats， and the time differences reaching the same water content gradually expanded as the dehydration proceeded； thus it won more time for moss plants in habitat E to carry out effective photosynthesis（here the relative water content is not less than 35%） and physiological and structural adjustment during the later dehydration. In total， G.pilifera plants in exposed rocks represented much stronger water stress tolerance than that in shaded rocks， but the later was able to increase the external water to make up the deficiency of rapid water loss. This might be the trade-off strategy of water absorption and retention for G.pilifera lived in different micro-habitats.

    Spatial Distribution Characteristics of All Genera in Rhamnaceae of China
    Ya-Li LI, Li-Chuan QIU, Jian WANG
    2021, 41(2):  191-198.  doi:10.7525/j.issn.1673-5102.2021.02.006
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    Spatial distribution characteristics of all genera in Rhamnaceae of China are based on the distribution data of species of Rhamnaceae in China and county of China. Spatial distribution data of genera were made in ArcGIS software. Area， perimeter and number of every patch were gotten from the maps. Simpson’s evenness indexes， shape indexes and largest patch indexes were calculated. Spatial distribution characteristics of all genera in Rhamnaceae can be analyzed. The experiment was conducted to determine the minimum range of spatial distribution， quantitative research， plant protection， utilization， resource development and restoration. Results showed that： ①Alphitonia， Scutia， Chaydaia， Berchemiella， Colubrina， Gouania and Ventilago arestenochoric genera. Paliurus， Rhamnella， Sageretia， Hovenia， Berchemia， Ziziphus and Rhamnus areeurychoric genera. Both the stenochoric genera and eurychoric genera account for 50% of the total genera of Rhamnaceae in China. The total area of stenochoric genera is less than 30×104 km2， Berchemiella mainlydistribute in the central subtropical zone of China. Other stenochoric genera mainly distribute near the tropic of cancer and south of it. The total area of eurychoric genera is greater than 40×104 km2.. They are distributed in most parts of our country， and scattered in the northwest.②Rhamnus has the largest distribution range and total area in Rhamnaceae of China， which almost covers the distribution range of all other genera. It can reflect the spatial distribution and characteristics of Rhamnaceae in China. ③Alphitonia is the only genus of continuous and the narrowest distribution of Rhamnaceae in China， and other genera are disjunction. Chaydaia， Gouania， Berchemia and Rhamnus only have the main distribution area. Scutia， Sageretia and Hovenia have main and secondary area. Berchemiella， Colubrina， Ventilago， Paliurus， Rhamnella and Ziziphus are all scattered area.

    Observation and Comparison of Root Primary Xylem Archy in Different Chlorophytum Varieties
    Xue-Peng FU, Le WANG, Jia-Le WEN, Kai-Fang YANG, Xiao-Jie YANG
    2021, 41(2):  199-204.  doi:10.7525/j.issn.1673-5102.2021.02.007
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    Three Chlorophytumcomosum varieties were used to investigate the relationship between the root primary xylem archy and C.comosum varieties， diameter of the roots and living conditions by using sections cut free-hand and stained with Safranin. The root primary xylem archy in the roots of different varieties were different. The root primary xylem archy in the roots of C.comosum in soil culture were also different from those under hydroponic condition， and the number of xylem bundles in the roots of C.comosum under hydroponic conditions was much less than that in soil culture. Under hydroponic conditions， there was a positive correlation between the number of xylem bundles in the roots of C.comosum and the diameter of the roots， however， the root xylem archy in C.comosum ‘Yinxin’ and ‘Yinbian’ has no correlation with the roots diameter of C.comosum.

    Photosynthetic and Stomatal Characteristics of Pinus koraiensis and P.sibirica under Low Temperature Stress
    Fang WANG, Zhi-Min LU, Jun WANG, Shi-Kai ZHANG, Yu-Xi LI, Shao-Chen LI, Jian-Qiu ZHANG, Yu-Chun YANG
    2021, 41(2):  205-212.  doi:10.7525/j.issn.1673-5102.2021.02.008
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    Both Pinus koraiensis and P.sibirica with strong cold resistance are famous forest species in the cold temperate zone. Compared with P.koraiensis， P.sibirica has stronger cold resistance. In order to explore the physiological response and cold resistance mechanism of the two species under low temperature stress，5-year-old P.koraiensis and P.sibirica seedlings were used to be conducted by low temperature treatment. By setting three stress temperatures（0℃， -20℃ and -40℃） and three stress times（6， 24 and 48 h）， taking 20℃ as control， which were used to study the photosynthetic characteristics and stomatal characteristics of P.koraiensis and P. sibirica under low temperature stress. By t test and variance analysis， there were significant differences （P<0.05） between P.koraiensis and P.sibirica for each photosynthetic index and stomatal density， both low temperature and low temperature stress time had an extremely significant effect on each photosynthetic index of P.koraiensis and P.sibirica（P<0.01）， and low temperature also had an extremely significant effect on stomatal opening size and stomatal area of P.koraiensis and P.sibirica（P<0.01）. The net photosynthetic rate， stomatal conductance and transpiration rate in P.koraiensis were significantly higher than those in P.sibirica at 20℃（without stress） and 0℃（low temperature stress）. However， under -20℃ for 6 h， each photosynthetic index in P.sibirica was higher than that in P.koraiensis. With the decrease of temperature and the extension of stress time， each photosynthetic index of the two species showed a decreasing trend. The stomatal density in P.koraiensis was significantly higher than that in P.sibirica. Before stress（20℃）， the stomas of P.koraiensis and P.sibirica were all oval. The stomatal opening size and stomatal area of the two species decreased significantly with the decrease of temperature.

    Variation of Flower Phenotypic Traits on 195 Syringa oblata SeedSeedings Single Trees
    Rui-Qing HE, Yun-Yao YANG, Zeng-Hui HU, Jing WU, Ping-Sheng LENG
    2021, 41(2):  213-220.  doi:10.7525/j.issn.1673-5102.2021.02.009
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    To accurately grasp the variation degree of floral traits and the phenotypic diversity level of Syringa oblata， so as to provide a theoretical basis for the breeding of S.oblata excellent varieties. The 195 seed seedlings of S.oblata trees around 8 years old were used to analyze the genetic variation， trait correlation and clustering relationship of 5 quantitative traits and four quality traits. Five quantitative traits were extremely significant differences among the 195 trees（P<0.01）， and accorded with the normal distribution. The genetic variation coefficients were from 19.72%-30.22%， with an average of 24.97%， and the inflorescence length was the largest. According to the ISCC-NBS method， the group colors were divided into six color groups， namely white， pale pink， purple， purplish red， purplish pink and violet groups. Purple group（54.35%） and purplish red group（32.31%） accounted for the largest proportion； the diversity index of four quality traits ranged from 0.36 to 1.09， with an average of 0.86， of which the corolla lobes（state） were the largest， indicating that the quality traits were abundant within 195 trees. By correlation analysis， there were significant positive correlations between the five quantitative traits， and the correlation coefficient between inflorescence length and inflorescence width was the largest（0.767）. By cluster analysis， the 195 samples of S.oblata can be divided into three categories based on euclidean distance. The first type was characterized by corolla tube length； the second group was characterized by long inflorescence； the last type was characterized by corolla lobes. The phenotypic variation of floral traits in the tested genetic resources of S.oblata was richer.

    Establishing an Efficient Regeneration Protocol in vitro for Rapid Micro-propagation of Two New Varieties of Ilex verticillata L.
    Rui LIU, Jia-Yi ZHANG, Chao-Qi GE, Lu-Lu XIE, Ying CHEN, Can ZHANG, Xi CHEN
    2021, 41(2):  221-231.  doi:10.7525/j.issn.1673-5102.2021.02.010
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    An efficient and rapid micro-propagation system using tissue culture for two new varieties “Red Sprite” and “Winter Gold” of Ilex verticillata L. was established with the young shoots as explants， and an optimum method produced large numbers of genetically uniform seedlings and maintained its excellent character for I.verticillata L was offered. The optimal medium for axillary bud induction was MS+1.0 mg·L-1 6-BA+0.2 mg·L-1 NAA， and the induction rate of both varieties were 100% respectively. The preferred medium for cluster bud proliferation was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+0.5 mg·L-1 CPPU. The highest proliferation coefficient was 5.7 for “Red Sprite”， and 4.5 for “Winter Gold” for 25 d culturing respectively. The optimal seedling medium for “Red Sprite” was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+1.0 mg·L-1 TDZ+1.0 mg·L-1 GA， and that of “Winter Gold” was MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA+0.05 mg·L-1 TDZ+1.0 mg·L-1 GA， the seedling height was up to 6.80 and 4.38 cm respectively. The optimal rooting induction medium was 1/2MS+0.4 mg·L-1 NAA+0.4 mg·L-1 IBA+1.0 mg?L-1 activated carbon， the rooting rate of “Red Sprite” and “Winter Gold” were 95.0%， or 97.0% respectively. The best transplanting substrates were peat∶perlite=3∶2， with a survival rate of 95.7% for “Red Sprite” and 96.8% for “Winter Gold” respectively.

    Dynamic Changes of Endogenous Hormones in Rootstocks and Scions within Nurse Seedling Graft in Camellia oleifera Under Wound
    Wei LONG, Xiao-Hua YAO, Le-Yan LÜ
    2021, 41(2):  232-242.  doi:10.7525/j.issn.1673-5102.2021.02.011
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    In order to analyze the physiological response to the surface healing of rootstocks and scions， and to provide the theoretical support for the growth mechanism， the dynamic changes of endogenous hormones in post-traumatic stress response in Camellia oleifera A. under wound were studied. The six-year-old Changlin 18 and Changlin 53 trees were used， and the contents of IAA， ABA， TZR， Zeatin， SA， JA were assayed using HPLC-MS at different wound stages， and the changes of endogenous hormone in different period and relationship between varieties were analyzed. The results showed that the content of IAA， SA and JA decreased with the increasing time of wound in scions of Changlin 18， and the content of TZR and Zeatin increased first and decreased later which they were respectively reached the highest in S10 and S0. The content of ABA rose continually and reached the peak in S10. In scions of Changlin 53， the contents of IAA and JA would decrease after peak； the TZR， Zeatin， SA were gradually increased after minimum. The content of ABA was gradually decreased in S0 after maximum. The contents of endogenous hormones were higher in Changlin18 than in Changlin53 in stem of rootstock between varieties. The endogenous hormones were decreased in Z0 and increased in Z10 in varieties except the contents of ABA and SA in Changlin53 increased first and then decreased. In stem and root of rootstocks， the contents of endogenous hormones were higher in Changlin 18 than in Changlin 53 except JA； The contents of IAA and ABA in stem of rootstocks were higher than roots which other endogenous hormones in root of rootstocks were higher than in stem. There was difference in the ratio of hormone in varieties and part of rootstocks. The ratios of IAA/ABA， IAA/TZR， IAA/Zeatin， IAA/JA， ABA/TZR， ABA/Zeatin， ABA/JA in scions and rootstocks were higher in Changlin 53 than in Changlin 18. The ratio of SA/IAA in scions was higher in Changlin 18 than Changlin 53， and it was higher in Changlin 18 than Changlin 53 in ratios of SA/JA， SA/ABA of scions while it was higher in Changlin 18 in stem of rootstocks. The ratio of TZR/SA， TZR/JA in scions and stem of rootstocks were higher in Changlin 18 than Changlin 53. There was the significantly positive correlation in IAA and JA after wound， and the correlation of IAA and SA， SA and JA had difference between varieties， the significantly positive correlation in Changlin 18 and the significantly or significantly negative correlation in Changlin 53. There was the significantly positive correlation among TZR， Zeatin， SA and the significantly negative correlation among TZR， Zeatin， SA. In stem of rootstocks， there was very significantly positive correlation among hormones in Changlin 18， and there was the significantly positive correlation among TZR， IAA and JA， Zeatin and IAA and JA while there was the significantly negative correlation in TZR and SA， Zeatin and SA. There was significantly negative correlation in SA and JA. In stem and root of rootstocks， there was the difference in correlation of SA and other hormones among varieties， and other hormones in Changlin 18 and Changlin 53 had the same significantly positive or negative correlation. To sum up， it was easy to callus formation in surface of wound in Changlin 18， but it was difficult in Changlin 53 because there was much more high-level hormone of stress-resistant so that it would be possible that affects healing after grafting wound 10 min. The differences in ratio and contents of hormones between rootstocks and scions may be affected by the morphological reconstruction of grafted unions and the growth of scions in later stage.

    Cloning and Sequence Analysis of Squalene Synthase(SQS) Gene from Dioscorea bulbifera L. Microtuber
    Wei-Zhong KE, Wen-Juan ZHONG, Kai-Ying LIU, Xiang-Yuan LI, Ming-Hua YIN
    2021, 41(2):  243-250.  doi:10.7525/j.issn.1673-5102.2021.02.012
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    The core fragment of SQS gene was screened from the transcriptome database of Dioscorea bulbifera L. microtuber， the conservative fragment of SQS gene was obtained by RT-PCR， and the 3' and 5' terminal sequences of SQS gene were obtained by race technique， and the sequence was analyzed by bioinformatics. The SQS gene of D.bulbifera L. microtuber was 1 548 bp long and 415 bp amino acid sequence， with a theoretical molecular weight of 46 786.38 D and an isoelectric point of 5.97. SQS protein is a hydrophobic protein without signal peptide. It contains the functional domain necessary for SQS belonging to Isoprenoid_Biosym_C1 superfamily. The homology of SQS protein between D.bulbifera and other plants is high， and the amino acid similarity of SQS protein between D.bulbifera and D.zingiberensis is 96.4%. The full-length sequence of SQS gene was obtained for the first time from D.bulbifera L. microtuber， which has the typical characteristics of SQS homologous gene. It provides the basis for further study on SQS gene structure， gene expression and gene mutation of D.bulbifera microtuber， and provides data support for the correlation analysis of positive selection site and function of SQS， a key enzyme of triterpene synthesis pathway in Dioscorea.

    Expression Profile Analysis ofXylem Development Regulated by BpNAC012 Gene from Betula platyphylla
    Yi-Ping GUO, Jia-Xin LIU, Ying YU, Chao WANG, Chuan-Ping YANG
    2021, 41(2):  251-261.  doi:10.7525/j.issn.1673-5102.2021.02.013
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    NAC transcription factors have been considered to be the switches that regulate the formation of plant secondary cell walls. Our previous work indicated that the gene BpNAC012 was involved in the secondary cell wall biosynthesis of birch. In order to identify the downstream target genes regulated by BpNAC012， transcriptome sequencing of stems from BpNAC012over-expression and RNAi plants was conducted， and the differentially expressed genes were identified compare to the wild type birch. The results showed that， in contrast to the wild type plants， 627 genes were up-regulated and 229 genes were down-regulated in the over-expression transgenic line， and 299 up-regulated genes and 207 down-regulated genes were identified in the RNAi lines. The expression changes of downstream genes in the overexpression transgenic lines were much greater than that in the RNAi plants. Inhibition of BpNAC012 expression significantly affected the expression of genes related to protein modification and transport. The differentially expressed genes were enriched in pathways such as receptor signaling pathways， nutritional metabolism， amino acid synthesis， and phenylpropane biosynthesis. In addition， BpNAC012 can regulate the expression of genes related to cellulose， lignin bio-synthesis and xylem development， as well as a variety of transcription factors. This study laid the foundation for further analysis of the molecular regulatory mechanism of BpNAC012 in secondary cell wall bio-synthesis of birch.

    Effects of Different Canopy Density on the Chlorophyll Fluorescence and Water Storage Capacity of Sphagnum palustre in the Edge of the Tuntianjing Sinkhole in Bijie City
    Kuang-Zheng SHI, Zhao-Hui ZHANG, Chun-Mei HE, Zhi-Hui WANG
    2021, 41(2):  262-269.  doi:10.7525/j.issn.1673-5102.2021.02.014
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    In order to enrich the understanding of the growth environment of Sphagnum， explore its possible impact on the formation of erosional karst tiankeng， and find more suitable growth conditions for it， we used gradient method， with S.palustre of different canopy density environments in the edge of the Tuntianjing sinkhole in Bijie City， to definite the chlorophyll fluorescence parameters and water storage capacity. Results showed： ①The initial fluorescence（Fo） and maximum fluorescence（Fm） of S.palustre increased significantly with increasing canopy density（Fo：224.67-264； Fm：716-785.33）. The maximum photochemical efficiency（Fv/Fm） of PSⅡ decreased firstly and then became constant； photochemical quenching coefficient（qP） and photosynthetic electron transport rate（ETR） increased significantly（qP：0.17 -0.46； ETR：5.72-9.68） with increasing canopy density； non-photochemical quenching coefficient（NPQ） decreased significantly from 1.57-0.8. ②With canopy density increased， the rate of water absorption and water storage capacity of S.palustre were significantly increased， the natural water absorption and the saturated water absorption increased from 14.36-26.37 and from 21.7-27.01， respectively； the natural water storage increased from 3 032.97-12 816.01， and the saturated water storage increased from 4 576.34-13 175.97. ③Although S.palustre has a good sunproof， they are more suitable for living in the low light conditions. As canopy density increased， it was beneficial to accumulate chlorophyll， promote photosynthesis and increase the accumulation of biomass， leading to a stronger water storage capacity. Competition can inhibit the growth of S.palustre， share soil moisture， and a certain moisture was very important for the growth of S.palustre. ④The good water-holding properties and water absorption of S.palustre have an important role in slowing surface runoff that slowing down in geological processes. The difference in water holding capacity of S.palustre under different canopy densities can affect the distribution of water flow to a certain extent， thus affecting the formation of erosion-type sinkholes.

    Respond of Soil Nutrient to Plowing and Reseeding of Achnatherum splendens in the Qinghai Lake Region
    Tao LIU, Di ZHU, Ting LÜ, Rui-Fang LIANG, Feng LIU, Zi-Lan MA, Yi-Kang LI, Yu-Zhi MA, Xu SU
    2021, 41(2):  270-280.  doi:10.7525/j.issn.1673-5102.2021.02.015
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    In order to explore the vertical variation character of soil nutrients caused by plowing and reseeding treatments， the grassland of Achnatherum splendens was used to analyze the effects of two restoration with plowing in 1958 or reseeding in 1992 nutritional distribution characteristics of 0?10， 10?20， 20?30， 30?40 and 40?60 cm soil in the zone of the Qinghai Lake of the Qinghai-Tibet Plateau. The results showed that the soil organic carbon and the content of total K， the plot under plowing and reseeding disturbance， were significantly higher than natural A.splendens grassland. Both disturbances were beneficial to the shallow layers of soil to enrich available phosphorus and organic carbon nutrients， not to the restoration of total N（P<0.05）. The contents of available K， total N and organic carbon decreased as the depth of the soil increases， but the soil bulk density， soil pH， the content of total P and K were not significantly different among each layers of plowing soil. Compared with the natural group， the content of available N was significantly decreased at each layers of reseeding soil（P<0.01）， but both disturbances caused significantly higher total K content， and significantly lower total N content in the soil 40?60 cm（P<0.05）. Compared with the natural group， there were significantly positive correlation between the contents of total N and K（P<0.05）， not significant correlation between the contents of available P and N after reseeding， both disturbances aroused the negative correlation between the soil nutrients and the soil bulk density. The pH values of the soil decreased significantly after plowing and reseeding， and plowing treatment could concentrate the soil available nutrients in the surface layer. The reseeding treatment accelerated the turnover of soil nutrients， and the increase of output promoted the restoration of aboveground vegetation， Beside the total potassium content. Both disturbances caused the recovery of total nutrients in different soil layers will be an extremely slow process.

    Leaf Phenotype Variation and Heterosis in F1 Progeny of Cross Between Castanea mollissima and C.henryi
    Ping-Sheng ZHANG, Xi-Bing JIANG, Bang-Chu GONG, Yang XU, Jun-sheng LAI, Cong-Lian WU
    2021, 41(2):  281-294.  doi:10.7525/j.issn.1673-5102.2021.02.016
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    In order to explore the genetic variation and heterosis of leaf traits in cross F1 progeny of Castanea， 14 traits of leaf phenotype and photosynthetic physiology of F1 progeny（including 235 individuals） of 9 combinations （including interspecific and intraspecific cross between C.mollissima and C.henryi） and their parents were studied by using the statistical methods of Nested anova， Pearson correlation analysis and principal component analysis. The results showed： ①The average coefficient of variation of 14 traits ranged from 4.40%-27.76%， indicating that there were abundant genetic diversity in leaf phenotype and photosynthetic physiological traits in F1 hybrid progeny of Castanea. ②The average of phenotypic differentiation coefficient among combination were 32.32%， which showed that the variation within combinations was the main source of variation. ③The mid-parent heterosis rate of 14 traits ranged from -27.16%-90.53%， and the heterosis of different combinations was different. ④The Pearson correlation analysis showed that there were 46 and 13 pairs of correlations reached extremely significant levels（P<0.01） and significant levels（P<0.05）， respectively， and 5 leaf phenotype traits， such as leaf length and leaf width， and 4 photosynthetic physiological indexes， such as chlorophyll a and chlorophyll b， had a great influence on each other. ⑤2 crossing combinations ‘Kuili’×YLZ No.15 and ‘Kuili’×YLZ No.1 were finally selected by principal component analysis， and their offspring had the characteristics of large leaf area， high dry matter and photosynthetic pigment content， which can provide the main materials for further screening of excellent offspring.

    Cloning and Expression Analysis of Two Key Genes of Jasmonic Acid Synthesis in Response to Endophytic Infection from Rehmannia glutinosa
    Shu-Ping PENG, Cheng-Ming DONG, Yun-Hao ZHU
    2021, 41(2):  294-301.  doi:10.7525/j.issn.1673-5102.2021.02.017
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    Allene oxide synthase（AOS） and 12-oxophytodienoate reductase（OPR） were the key genes of jasmonic acid biosynthesis in the plants. The genes of AOS and OPR responding to endophytic fungus infection were screened from the interaction transcriptome of Rehmannia glutinosa with endophytic fungus GG22. Specific primers were designed to the open reading frame of RgAOS and RgOPR. Bioinformatics analysis of the sequences and theirs encoded product were performed， and real-time fluorescent quantitative PCR was used to analyze the expression patterns of RgAOS and RgOPR in different tissues infection by endophytic fungus GG22. The open reading frame of RgAOS was 1 626 bp， encoding 541 amino acids with a molecular weight of 60.2 kD. The open reading frame of RgOPR was 1 197 bp， encoding 398 amino acids with a molecular weight of 44.07 kD. QPCR analysis showed that the expression of RgAOS was the highest in roots， the lowest in flowers， and the expression of RgOPR was higher in flowers and leaves， respectively. Endophytic fungus GG22 induced the expression of RgAOS and RgOPR in R.glutinosa. The RgAOS and RgOPR were successfully cloned from R.glutinosa， laying the foundation for further study on the biological activity of Jasmonic acid substances in R.glutinosa and claiming the molecular mechanism of endophytic fungi in the regulation of secondary metabolites of R.glutinosa.

    Effect of Exogenous ATP on Cold Tolerance of Brassica campestris Seedlings
    Xin-Dan HU, Yao LI, Xiao-Hua ZHANG, Juan-Hong LIANG, Teng-Guo ZHANG
    2021, 41(2):  302-311.  doi:10.7525/j.issn.1673-5102.2021.02.018
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    The effects of exogenous ATP on the cold tolerance of rape seedlings were studied with “Longyou 7”Brassica campestris L. as experimental material. Compared with low temperature stress alone， MDA content and content of rapeseed seedlings were significantly decreased after exogenous ATP pretreatment and low temperature stress， while chlorophyll content and antioxidant enzyme activities（SOD， POD， CAT， APX） were observed. The expressions of RBOHD， RBOHF， CPK4 and CPK5 were increased. Compared with exogenous ATP+ low temperature， MDA content was significantly increased under EGTA+exogenous ATP+ low temperature treatment. Total chlorophyll content， T-AOC enzyme activity， RBOHD and RBOHF gene expression were all increased. Significantly decreased， MDA content was increased significantly under DMTU+exogenous ATP+low temperature treatment， total chlorophyll content， Ca2+-ATPase activity， CPK4， CPK5 gene expression were decreased， indicating that exogenous ATP affected cold tolerance of rape seedlings by Ca2+ and H2O2-dependent mechanisms.

    Establishment and Optimization of SSR-PCR Reaction System for Casuarina
    Zhen LI, Chong-Lu ZHONG, Yong ZHANG, Yong-Cheng WEI, Jing-Xiang MENG
    2021, 41(2):  312-320.  doi:10.7525/j.issn.1673-5102.2021.02.019
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    In order to obtain the optimal reaction medium of SSR-PCR from casuarina， 24 somaclonal clones from 4 species including Casuarina equisetifolia， C.junghuhniana， C.glauca and C.cunninghamiana， were selected as experimental materials. L16（45） orthogonal design was used to determine the optimum concentrations of four factors（primer， Mg2+， dNTP， Taq polymerase） within four concentration levels in the SSR-PCR reaction system of Casuarina respectively， and visual analysis and analysis of variance were used as evaluation. The results showed as follows： The three factors（primer， Mg2+， dNTP） had significant differences on the SSR-PCR reaction systems（P＜0.01） respectively， and the order of significance was primer＞Mg2+＞dNTP， while Taq polymerase had no significant difference. The two optimal SSR-PCR reaction systems（10 μL） of Casuarina were determined， including system 6， which consisted of 1×PCR buffer， 2ng template DNA， 0.5 μmol·L-1 primer， 1.5 mmol·L-1 Mg2+， 0.1mmol·L-1 dNTP， 0.5 U Taq polymerase， and system 15， which contained 1×PCR buffer， 2 ng template DNA， 0.5 μmol·L-1 primer， 1.75 mmol·L-1 Mg2+， 0.2 mmol·L-1 dNTP， 1.25 U Taq polymerase. The system 6 was better than system 15. 60℃ was determined as the optimal annealing temperature of the fluorescent primers M26 and M36.