Table of Content

15 March 2015, Volume 35 Issue 2

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Characteristics of Leaf Epidermis of Pseudostellaria(Caryophyllaceae) in China

LI Chao；GE Wei-Na；LI Ming

2015, 35(2):  161-164.  doi:10.7525/j.issn.1673-5102.2015.02.001

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We observed the leaf epidermis characteristics of six species from the genus Pseudostellaria in China under the light microscopy, measured and calculated the size and density of epidermal cell and stomata apparatus. The shapes of leaf epidermal cells are irregular, anticlines walls are repand or sinuate. The size of adaxial epidermis cell is about 61.7×95.8 μm-115.3×179.3 μm, the density is about 76-158 mm^-2, the size of abaxial epidermis cell is about 46.4×72.4 μm-91.9×150.4 μm, and the density is about 104-287 mm^-2. Most of species have stomata apparatus only on abaxial epidermis, the type of stomata apparatus is anomocytic, the density is about 30-143 mm^-2, and the size is about 21.5×28.6 μm-30.4×47.2 μm. The stomata index is about 19.5-31.6. Stomata is oblong and thickened at the ending parts. Pseudostellaria is related to Stellaria, and is a derived taxon.

Observation Process of Free Nuclei Division and Cellularization in Ginkgo biloba L. Endosperm

LU Zhao-Geng；LUO Kai-Ge；LI Wei-Xing；PAN Ye；ZHANG Min；WANG Li*

2015, 35(2):  165-171.  doi:10.7525/j.issn.1673-5102.2015.02.002

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Using slicing and total dissection techniques, we observed the process of division and proliferation of free nuclei, and endosperm cellularization in Ginkgo biloba L. We analyzed the proliferate development of endosperm cells by the means of disassembling endosperm with the cellulose enzymes. The results showed that the division and proliferation of free nuclei occurred 5-30 days after pollination and cellularization occurred about 30-65 days after pollination.The female gametophyte mitosis resulted in many free nuclei, which was spherical shape with the average diameter of 2-4 μm. At the initial stages, the free nuclei distributed in the central cytoplasm. However, with the volume of central vacuoles increasing, cytoplasm and nuclei were pushed to around. Then, one layer of opening cells that had no cell wall at the side near central vacuole formed in the periphery after the formation of about 4 000-5 000 nuclei. Cells around grew centripetal and dissociative, till cavum was filled with endosperm cells and the cellularization was finished. The proliferate development of the endosperm cells took on “S”-shape curve, which was slowly at the early stage after pollination, then it changed faster during 40-55 days after pollination, and the number of endosperm cells reached the maximum at 65 days after pollination. Afterwards, the number of endosperm cells did not significantly changed. Therefore, the division and proliferation of free nuclei have unique traits, and the number of endosperm cells play a significantly role in the development of seeds in G.biloba.

Micromorphological Observations on Leaf Epidermis and Pollen of Prinsepia Royle in China

YANG Rui-Lin1；WEI Xue-Zhi2*

2015, 35(2):  172-178.  doi:10.7525/j.issn.1673-5102.2015.02.003

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We examined the micromorphology of leaf epidermis and pollen characteristics of four species of Prinsepia Royle in China by using scanning electron microscopy. Among them, it was the first time to report these above information of P.sinensis and P.utilis in Taiwan. The pollen grains were subglobose, radial symmetry, and tricolporate, the exine sculpturing was stripe with equatorial aperture, and the range of value of P/E was from 0.91 to 0.97. The size range varied within (19.62-27.30)μm×(21.53-29.59) μm. There were two types of pollen grains which were blunt- triangle and trifid round in polar view, two groups of aperture were goniotreme and peritreme, pollen had exine with perforations and exine with no pores. Two groups of pollen grains were mucus drip on the pollen exine without mucus drip. There were more differences in the density and spacing of vallate as well as the characters of colpi. Four species were divided into two types, epidermal hair or not. There were large differences between three species including the characters of cuticle, stoma density, waxy and stoma form. We provided the references about palynology and the micromorphology of leaf epidermis for classification study of native Prinsepia Royle, and theoretic basis for introduction and cultivation.

Tissue Culture System of Axillary Proliferation of Acacia crassicarpa

HU Feng1,2；SHI Qiong1；HUANG Lie-Jian1*

2015, 35(2):  179-184.  doi:10.7525/j.issn.1673-5102.2015.02.004

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We studied the tissue culture system of Acacia crassicarpa by using the stemsegment with buds collected from 16-years plant as explants. The buds could be induced successfully on 2% of MS+sucrose with 67.33% of the survival rate and 80.56% of shooting rate after sterilized with 75% of alcoho1 and 0.1% of mercuric chloride for 30 s and 12 min, respectively. Low concentration of activated carbon effectively alleviated the verification, the best medium for multiplication was Modified-MS+6-BA 0.5 mg·L^-1+NAA 0.25 mg·L^-1+Ac 0.1 g·L^-1+sucrose 3% and the multiplication number was 3.72 after 35 days. The best medium for rooting was Modified-MS+IBA 0.5 mg·L^-1+2% of sucrose, and the rooting rate was 98.83%. The survival rate reached 90.00% after transplanting seedlings to the nutrition cup with yellow soil.

In Vitro Culture and Plantlet Regeneration of Quercus shumardii

Lü Xiu-Li1,2；SHEN Lie-Ying1；SHI Ji-Sen2；ZHANG Chun-Ying1*

2015, 35(2):  185-190.  doi:10.7525/j.issn.1673-5102.2015.02.005

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With semi-lignified shoots sprouted from the girdling of 13 years old Quercus shumardii as explants, we established the regeneration system of Q.shumardii by in vitro culture. When the WPM was supplemented with 1.0 mg·L^-1 BA , 0.1 mg·L^-1 NAA and 5 mg·L^-1 Biotin, 5 mg·L^-1 acetylcholine, 5 mg·L^-1 vitamine B2, and cultured for 40 d, the multiplication coefficient was 2.15. The rooting rate of single bud was 10% when cultured in 1/2 WPM supplemented with 2.0 mg·L^-1 NAA, 5 mg·L^-1 Biotin, 5 mg·L^-1 acetylcholine, 5 mg·L^-1 vitamine B2. The rooting rate was about 80% when roots were induced out-tube. Our work established technical system for tissue culture and plantlet regeneration of Q.shumardii with a solid foundation for the extension of its good quality seedlings.

Elaiosome Development in Chelidonium majus L. Seeds

YANG Sen1；LI Jian-Xia1；XIA Xiao-Fei2；ZHAO Liang-Cheng1*

2015, 35(2):  191-199.  doi:10.7525/j.issn.1673-5102.2015.02.006

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We investigated the initiation, development and reserves of elaiosome in Chelidonium majus L.(Papaveraceae) seeds by parafin sectioning, SEM and histochemical tests. The elaiosome develops very early and its initial cells originate from the outmost layer cell of raphe at the stage of two to four nucleus embryo sac, so it belongs to the epidermal origin. The divisions of basal cells of elaiosome mainly take place before fertilization, and the growth and expansion of top cells mainly occur after fertilizing, leading to the remarkable enlargement of the elaiosome size. The initiation of the elaiosome has no relation with the ovule fertilization, but a normal growth of the fertilized ovule is needed for the later proper development of the elaiosome. The only reserve of the elaiosome in mature seeds is fat and the fat accumulation mainly occurs at the stage of near seed maturation. The elaiosome of Ch.majus should be referred to as strophiole in term of its origin position and its main function is to attract ants to disperse the seeds.

Photosynthesis and Antioxidant Defense Strategies in Overwintering Plants of Tsuga chinensis and Cyclobalanopsis stewardiana

ZHANG Qiang1,2；GUO Chuan-You1；ZHANG Xing-Wang1；BAI Kun-Dong2*；JIANG De-Bing3

2015, 35(2):  200-207.  doi:10.7525/j.issn.1673-5102.2015.02.007

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We selected two dominant tree species including Tsuga chinensis and Cyclobalanopsis stewardiana grown in the evergreen needle-leaved and broadleaved mixed forest at high-altitude on Mao’er Mountain, and studied their seasonal changes in photosynthetic pigments, net photosynthetic rate(A_max), chlorophyll fluorescence and antioxidative system. T.chinensis and C.stewardiana down-regulated the values of chlorophyll(Chl) content, predawn maximum photochemistry efficiency of PSⅡ(F_v/F_m-predawn), A_max and stomatal conductance(G_s) to the minimum during winter stress. The activities of superoxide dismutase(SOD), peroxidase(POD), catalase(CAT) and the contents of ascorbic acid(AsA), glutathione(GSH) and carotenoids(Car) of C.stewardiana were the highest in winter, which showed good synergy in resisting adverse external environments and scavenging reactive oxygen species(ROS), while only AsA, GSH and Car in T.chinensis maintained good synergy in resisting winter stress. The seasonal changes of MDA contents in T.chinensis and C.stewardiana were correlated with the seasonal variations of environment as well as the seasonal variations of antioxidative enzyme activities and antioxidants accumulation. Therefore, T.chinensis and C.stewardiana had different antioxidant-defense strategies, but the same photosynthetic adaptation strategy was down regulation for winter acclimation.

Salt Tolerance of Transgenic Tobacco Containing PsATX and PsSOD Genes of Polygonum sibiricum Laxm.

MA Jing1；QU Chun-Pu2；XU Zhi-Ru1,2*；YANG Chuan-Ping2；LIU Guan-Jun2

2015, 35(2):  208-217.  doi:10.7525/j.issn.1673-5102.2015.02.008

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We analyzed the salt tolerance of tobacco plants which expressed both PsATX and PsSOD genes of Polygonum sibiricum Laxm., constructed univalent expression vector pROKⅡ-PsATX, pROKⅡ-PsSOD and dual-gene expression vector pROKⅡ-PsATX-PsSOD respectively, and then transferred them into tobacco mediated by Agrobacterium tumefaciens. The transgenic tobacco were identified by PCR method. We measured the content of malondialdehyde(MDA), reactive oxygen, proline(Pro), and the activity of superoxide dismutase(SOD) in the transgenic tobacco under salt stress condition to identify the salt tolerance of these plants. The transgenic tobacco containing PsATX, PsSOD and PsATX+PsSOD genes respectively were obtained successfully. The integration of the foreign PsATX and PsSOD genes were confirmed by molecular identification. Under the treatment of 100 mmol·L^-1 NaCl solution, the content of MDA, reactive oxygen and proline in the co-transformed plants was lower than that of wild type and univalent transgenic plants, but the SOD activity were higher than them. By the salt test, the transgenic tobacco contained PsATX+PsSOD genes were more tolerant to salt stress than the transgenic tobacco contained PsATX or PsSOD genes.

Endogenous Hormone Contents of Cut Freesia Flowers during Vase Holding

LIU Ya-Jie；CHANG Ping；TANG Dong-Qin*

2015, 35(2):  218-224.  doi:10.7525/j.issn.1673-5102.2015.02.009

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We used the cut flowers of Freesia hybrida ‘Shangnong jinhuanghou’ to study the endogenous hormone content changes of petals in the process of flowering and aging during vase holding in distilled water. We determined the contents of auxin(IAA), gibberellic acid(GA), zeatin nucleoside(ZR) and abscisic acid(ABA) by enzyme-linked immunoassay, while measured ethylene releasing rate by gas chromatography, and observed the content changes of five endogenous hormones in petals during vase holding. The contents of endogenous ZR decreased constantly, and the lowest content decreased by 83.8% on the 10th day compared to that at beginning. The change trend of endogenous GA is similar to ZR content, the lowest content of GA was present on the 10th day, correspond to 3.8% by compared with that at beginning. However, the content of endogenous ABA, accumulating after a descend, dropped to the lowest level on the fourth day, equivalent to 52.4% of the content at beginning. Endogenous IAA content, showing a trend of decline after rising first, emerged a peak on the second day, and down to a minimum level on the 10th day, equivalent to 12.2% of the highest. At the same vase period, contents of the endogenous IAA, ZR and ABA had no significant difference in the different positions of inflorescence, but endogenous GA content in the upper floret of inflorescence is significantly higher than the base and mid in the first four days during vase holding. Change of ethylene releasing rate increased followed a constant decrease up to the end of vase holding. However, the ethylene releasing peak of mid position floret appeared on the second day, while the upper floret on the fourth day. The metabolic unbalance of endogenous hormone levels might be one of the causal factors of freesia flower senescence.

The Ecophysiological Response of Calligonum caput-medusae to Different Water Condition in Extremely Arid Region

HUANG Cai-Bian1,2；ZENG Fan-Jiang1,2；LEI Jia-Qiang1,2

2015, 35(2):  225-232.  doi:10.7525/j.issn.1673-5102.2015.02.010

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The pot experiment was conducted to study the changes of plant growth and physiological characteristics of 1- and 3-year-old Calligonum caput-medusae under different irrigation treatments (30%, 40%, 50%, 60%, and 85% field water capacity(FC)), and find out the suitable water condition for the plant growth. The plant height, canopy, base diameter, assimilating branch diameter, and the biomass of whole plant declined significantly with the decreasing of irrigation amount for both 1- and 3-year-old plants. At both growth stages, stem was the main organ for dry matter accumulation. The dry matter allocation proportion of stem was positively affected by the increasing irrigation amount, but that was negatively for root. However, C.caput-medusae of 3-year-old could not produce fruit at the lowest irrigation level, and the fruit yield was significantly lower at 85% FC than that by other treatments. The assimilating branches had high chlorophyll and MDA contents under lower irrigation treatment. The damage of plasmamembrane was serious under drought condition. However, there was a large number of proline accumulated in the assimilating branches to adapt to water stress. The damage of plasmamembrane was reduced with the increasing of irrigation, and the proline and soluble sugar accumulations were complemented each other. The chlorophyll content decreased significantly under the highest irrigation treatment for both 1- and 3-year-old C.caput-medusae. Based on all of the above determined indexes, the comprehensive adaptability was in the following order: 60% FC>85% FC>50% FC>40% FC>30% FC. Therefore,the medium irrigation level (60% FC) is appropriate for establishing C.caput-medusae seedlings in this area.

Effects of KCl on Active Oxygen Metabolism and Osmotica Accumulation in Avena nude L. Seedlings under NaCl Stress

LIU Jian-Xin；WANG Jin-Cheng；WANG Rui-Juan；JIA Hai-Yan

2015, 35(2):  233-239.  doi:10.7525/j.issn.1673-5102.2015.02.011

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We studied the effects of KCl on the plant growth, active oxygen metabolism and osmotica accumulation in Avena nude seedlings (‘Dingyou No.6’) under 120 mmol·L^-1 NaCl stress with sand cultivation in a greenhouse. Compared with 1 mmol·L^-1 KCl, 10 and 20 mmol·L^-1 KCl on A.nude seedlings, respectively, could significantly increase its weights of fresh and dry, relative water content, as well as its leaf activities of superoxide dismutase, peroxidase and ascorbate peroxidase, but decrease its the contents of O^·₂, H₂O₂, malondialdehyde and glutathione and membrane relative permeability under the NaCl stress. Moreover, the effect of 20 mmol·L^-1 KCl was higher than that of 10 mmol·L^-1, but there were no significant effects on the catalase activity and ascorbic acid content, however, the damage of salt stress was significantly increased when KCl concentration was up to 3.0 mmol·L^-1. The contents of soluble protein, soluble sugar and proline in A.nude leaves were significantly higher than that of others, when KCl concentration was 10 and 20 mmol·L^-1, but its free amino acids contents were significantly lower than that of 1 mmol·L^-1 KCl, and little variation in content of organic acids. The K⁺/Na⁺ ratio in leaves of A.nude seedlings under NaCl stress increased significantly, and the plasma membrane H⁺-ATPase activities demonstrated a hump-shaped curve with the increase of KCl. Therefore, the ratio 6∶1 of Na⁺/K⁺ could alleviate the negative impact of salt stress on A.nude seedlings through improving the ability of scavenging reactive oxygen species and osmotica accumulation.

Cloning and Expression Analysis of a Transcription Factor SlYABBY5 in Solanum lycopersicum

WANG Su-Su1；CHEN Guo-Ping1；ZHANG Yan-Jie1；YIN Wen-Cheng1；ZHU Ming-Ku1； ZHANG Jian-Ling1；TANG Qu-Lin2；HU Zong-Li1*

2015, 35(2):  240-249.  doi:10.7525/j.issn.1673-5102.2015.02.012

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YABBY family is a small plant-specific gene family, with a typically N-terminal C2C2-type zinc finger domain and the C-terminal helix-loop-helix YABBY domain. By homologous sequence analysis, a YABBY transcription factor was cloned from tomato (Solanum lycopersicon), and named SlYABBY5(GenBank accession number: AK246138). By bioinformatics analysis, the length of SlYABBY5 ORF is 549 bp, encoding 182 amino acid residues with relative molecular weight of 20.58 kD and an isoelectric point of 8.78. By real-time PCR analysis, SlYABBY5 gene might be involved in the development of tomato fruits and leaves. Expression of SlYABBY5 down-regulated under the treatments of IAA, ABA, GA₃, ASA and low temperature stress, while up-regulated under salt stress and leaves wounding. Our results suggest that SlYABBY5 might participate in the growth and development of tomato and abiotic stress responses, and provide the basis for unrevealing the function of SlYABBY5 gene in tomato growth and development as well as stress signaling pathway.

Cloning and Sequence Analysis of HRD Transcription Factors Gene from Two Kinds of Ephemeral Plants

CHEN Qin1,2；QU Yan-Ying1,2；LIU Jing-Jing1,2；ZHU Yan-Fei1,2；YAO Zheng-Pei1,2；CHEN Quan-Jia1,2*

2015, 35(2):  250-258.  doi:10.7525/j.issn.1673-5102.2015.02.013

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We isolated two transcription factor genes, named as CpHRD and SaHRD, from two kinds of ephemeral Conringia planisiliqua L. and Sisybrium altissimum L. by RT-PCR, respectively. By bioinformatics, both of CpHRD and SaHRD are possessing an intact complete open reading frame of 564 bp and sharing 88%, 87%, 87% and 86% of sequence identity with Eutrema salsugineum, Arabidopsis lyrata, Capsella rubella and Arabidopsis thaliana, respectively. The deduced polypeptide sequence of CpHRD and SaHRD are 187 amino acid residues with a predicted molecular weight of 20 kD with secondary protein structure of the polypeptide chains contain including 2 β-sheets, 5 α-helixs and random coils, belonging to hydrophilicity and stable of transmembrane protein. By phylogenetic tree analysis, CpHRD and SaHRD mainly exist in the nucleuscontains, and have a typical AP2/EREBP domain function structure, which may be involved in plant stresses response process of ABA, drought and low temperature. Our results can provide useful information about the molecular mechanism of HRD genetic mechanism of drought resistance, and will contribute a valuable resource for development and utilization of germplasm resources of ephemeral plant.

OsLOL2 Gene Clone of Rice and the Salt Resistance Analysis of Overexpression Arabidopsis thaliana

GUAN Qing-Jie1,2；WANG Zhen-Juan1；ZHENG Heng1；LIU Guang-Tao3；LIU Shen-Kui3

2015, 35(2):  259-269.  doi:10.7525/j.issn.1673-5102.2015.02.014

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A known LSD1-Like type zinc finger protein transcription factor OsLOL2 gene(LOC_Os12g41700) was cloned from the rice Longjing 11(Oryza sativa L. longjing11). By bioinformatics analysis, the length of the ORF of OsLOL2 gene is 519 bp, which encoded 172 amino acid residues, with 2 LSD1-Like zinc finger domain. The transformation of epidermal cells of onion by Gene gun shows OsLOL2 locates in the nucleus. By real-time quantitative PCR, the transcriptional level of OsLOL2 gene increased significantly in Longjing 11 roots and leaves induced under 30 mmol·L^-1 NaHCO₃, 80 mmol·L^-1 NaCl, and 10 mmol·L^-1 H₂O₂ stress condition. Overexpression of OsLOL2 gene in yeast showed resistance to saline growth. Under stress condition, by the germination rate analysis, the germination rate of transgenic Arabidopsis was higher than that of wild type under NaCl and H₂O₂ stress. The germination rate and resistance on germination late showed the growing of overexpression strain was better than WT on 1/2 MS plate which contained 100 mmol·L^-1 NaCl, 3 mmol·L^-1 NaHCO₃ and 2 mmol·L^-1 H₂O₂. OsLOL2 gene could improve resistance for plant salt alkaline, which was related to the ROS way of oxidative stress by salt-alkali stress. Our study can provide a theoretical basis for studying OsLOL2 gene involved in stress signal way and lay the foundation for creating of new rice with stress resistance.

Characterization of the Glycosyltransferase Gene UGT78H2 from Blackberry and Docking with Flavonoid Molecules

CHEN Qing1；JIANG Lei-Yu1；WANG Yan1；ZHANG Yun-Ting1；WANG Xiao-Rong2；TANG Hao-Ru1*

2015, 35(2):  270-278.  doi:10.7525/j.issn.1673-5102.2015.02.015

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Glycosylation plays a pivotal role in determining the diversities of plant small molecules. Glycosyltransferases are ubiquitous proteins involved in such reactions. However, the exact mechanisms of sugar donor or acceptor substrate recognition are still to be elusive. We reported the isolation of a new glycosyltransferase gene from blackberry plants(Rubus spp.), nominated as UGT78H2(GenBank Accession No.:KM061379) by the international UDP Glucosyltransferase Nomenclature Committee. The full length of the gene contained an open reading frame of 1 380 nucleotides, encoding a 50.6 kD polypeptides of 459 amino acids. By further analysis, the deduced enzyme was a typical family-1 glucosyltransferase with the plant secondary product glucosyltransferase signature motif in the C terminal. Phylogenetic trees revealed the clustering of UGT78H2 in the F node together with other Arabidopsis thaliana GTs, inferring the possible catalysis activity toward flavonoid compounds. Amino acids of UGT78H2 shared most, but only 88% identity to the RhUF3GT(UGT78H1) in Rose hybrid when aligned to other glycosyltransferases. Most abundant transcripts of the gene were observed in the immature blackberry fruits, a stage when little anthocyanins were produced. Additionally, transcription levels decreased dramatically with the maturation of fruits. We used the Medicago truncatula UGT78G1(PDB: 3HBF) as a template to homologically model the protein of UGT78H2 that was subsequently docked with 12 common flavonoid molecules. UGT78H2 was predicted to be a glucosyltransferase having maximum binding affinity towards quercetin, tangeritin and epicatechin.

Effect of Fire Disturbances on Short-term Soil Respiration and Its Components of Larix gmelinii and Betula platyphylla Forests in Xiaoxing’an Mountains

HU Hai-Qing1；WU Wei1；YUE Cai-Ling2；CHEN Wan-Ting2；ZHANG Ran1；LI Ying1；SUN Long1*

2015, 35(2):  279-288.  doi:10.7525/j.issn.1673-5102.2015.02.016

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In burned areas of Larix gmelinii and Betula platyphylla forests in Xiaoxing’an Mountains, with the unburned nearby areas as the control, we examined the rate of soil respiration in burned and control areas, and studied the differences of short-term soil respiration between the burned and control areas. By comparing the rate of soil respiration between the burned and control areas of two forest types in same months, there was significant difference in august(P<0.05), while it was not obvious in other months(P>0.05). The overall mean rate of soil respiration in B.platyphylla forest had a descending trend after fire disturbance, while it was increased after the fire in L.gmelinii forest. The specific performances were as follow: the mean rate of R_s over the growing season in the burned areas(3.91 μmol·m^-2·s^-1) was lower than the control areas(5.29 μmol·m^-2·s^-1) in B.platyphylla forest, and R_h, R_a, the overall mean rate of soil respiration over four months were also declined after the fire. The rate of R_s in burned plots(4.62 μmol·m^-2·s^-1) was higher than that in unburned plots(4.18 μmol·m^-2·s^-1) over the growing season in L.gmelinii forest, R_h, R_a and the overall mean soil respiration rate in burned plots was also higher than those in control plots. The variation in the soil respiration rate could be explained by soil water content and soil temperature and their interaction at the depth of 5 cm. By regression, fitted T, W and T×W model could explain that R_s rates in control plot of B.platyphylla and L.gmelinii forests were 82.9% and 88.6%, respectively. It was 88.8% in post-fire plot of B.platyphylla forest, higher than the control. While it declined to 72.9% in burned plot of L.gmelinii forest. We found out the change of fire disturbance on soil respiration, and would provide a scientific basic data on the effects of fire disturbance on short-term soil respiration in the soil respiration on the response of fire disturbance.

Effects of Trichoderma asperellum on the Physical and Chemical Properties and Nutrient Components of the Pot Coil Culturing Tissue-cultured Populus davidiana×P.bolleana Seedlings

Lü Man-Man1；LIU Zhi-Hua2；WANG Hui1；ZHU Guo-Dong1；YANG Xing-Tang1；ZHANG Rong-Shu1*

2015, 35(2):  289-296.  doi:10.7525/j.issn.1673-5102.2015.02.017

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In order to study the effect of Trichoderma asperellum ACCC30536 on the physical and chemical properties and nutrient components of the soil in a 60-day growing period under field conditions, we conducted pot culturing experiment of one-year-old tissue-cultured Populus davidiana×P.bolleana seedlings. Both soil in empty pots and soil in seedling-culturing pots were treated with T.asperellum conidia solution at the concentration of 5×10³ cfu·cm^-3 soil in the experiment (as T and T1 group), where bare pot soil and soil in a seedling-culturing pot without treatment were taken as control groups (Con and Con1, respectively). By ANOVA, the differences in inducing durations and levels of T.asperellum treatment significantly influenced the moisture content of the soil (P<0.05), while all treatments significantly decreased the moisture content (P<0.05). The differences in inducing durations and levels of T.asperellum treatment had no significant influence on the alteration of soil pH (P>0.05). Although T.asperellum treatment had a general buffering effect on soil pH, the pH was kept in the range of 6.84-6.97. The differences in inducing durations and levels of T.asperellum treatment significantly affected the content of organic matter, total nitrogen, total phosphorus, available phosphorus and total potassium (P<0.05), resulting in a highest organic matter content of 1.27 and 1.31 times of that of Con and Con1 respectively, among those of all sampling time points in all seedlingculturing pots (P<0.05), and a maximum total nitrogen content of 1.18, 1.18 and 1.25 times of those of Con, Con1 and T, separately (P<0.05), and a maximum total phosphorus content of 1.12 times of that of Con1(P<0.05), a maximum available phosphorus content of 1.13 times of that of Con1 (P<0.05), and a maximum total potassium content of 1.17 times of that of Con1 (P<0.05). Inducing duration had a significant influence on the available potassium content (P<0.05), while difference in treatment levels had no significant influence on the available potassium content of the soil (P>0.05). Furthermore, the interaction between the inducing durations and the difference in treatment levels of T.asperellum also significantly influenced the above indexes (P<0.05). Therefore, T.asperellum ACCC30536 conidia treatment can significantly affect the physical and chemical properties and the nutrient components of soil.

Effect of Thinning on Litter Decomposition and Soil of Two Stands in Changbai Mountain

LIANG Jing；WANG Qing-Cheng*；XU Li-Juan；WU Wen-Juan

2015, 35(2):  297-303.  doi:10.7525/j.issn.1673-5102.2015.02.018

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With poplar-birch stand and Larch-birch stand in Changbai Mountain, we studied two stands in three kinds of thinning intensity decomposition of forest litter accumulation and features, soil physical properties and change regularity of nutrient elements, and litter and soil carbon density and their relationships with the thinning intensity. Under different thinning intensity, there were significant differences in poplarbirch stand and larchbirch stand litter accumulation and decomposition rate. Litter nutrient content had a lower trend with the increase of thinning intensity. Litter carbon content increased with the increase of thinning intensity. Soil pH increased with the increase of thinning intensity obviously. Thinning affected the soil bulk density and the water content rate less. The soil carbon content reduced, and then increased with the increase of thinning intensity. The soil carbon density decreased with the increase of thinning intensity. Therefore, thinning is the main factors on litter decomposition and soil carbon accumulation.

Characteristics of Gap Makers and Plant Diversity in Picea asperata and Abies fabri Forest in Xiaoxing’an Mountains

WANG Ting1；DUAN Wen-Biao1*；CHEN Li-Xin1；RONG Li-Jie2；JING Xin1；WANG Jin-Ling1

2015, 35(2):  304-309.  doi:10.7525/j.issn.1673-5102.2015.02.019

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An investigation was conducted in a 1.5 hm² permanent standard plot of Picea asperata and Abies fabri forest to determine the species, basal diameter, DBH, height, breakage height, formation mode, decayed level of all gap makers, the species, abundance, height, cover of all herbs and shrubs, and calculate their important value and richness in June August, 2013. The number of uprooted trees was 36, the number of stem breakage and base breakage was 17 and 6, respectively. The value of Pielou evenness index(J), Simpson eco-dominance index(D), Shannon index(H) were listed in the order of all uprooted tree>stem breakage>base breakage. Within the plots of uprooted tree and breakage, the important value of Aegopodium alpestre was the biggest, 14.2 and 13.2, respectively. The dominant species in the plots of uprooted trees are Impatiens nolitangere and Urtica fissa, but Aconitum carmichaeli in the plots of breakage. Under the same decayed level of tree falls, species richness in the plots of uprooted trees was more than that in the plots of breakage. The herbs’ richness in the east, south, west, north plots of base breakage and stem breakage were smaller than that in the control, but opposite in uprooted tree plots.

Optimizing Sequence-Related Amplified Polymorphism Amplification System for Isatis indigotica Fort

WU Zheng-Qian1,2；LI Xue-Hu1*；LIANG Jian-Ping1；LU Xi-Hong1；XIN Zhi-Jun1；ZHOU Xiang1

2015, 35(2):  310-314.  doi:10.7525/j.issn.1673-5102.2015.02.020

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We established and optimized SRAP-PCR amplification system for Isatis indigotica Fort by the orthogonal design. The order of factors affecting the result of SRAP-PCR were Mg²⁺, dNTPs, primers, template DNA and Taq DNA polymerase. A suitable SRAP-PCR system for Chinese bayberry was that total 25 μL reaction system containing 1.5 mmol·L^-1 Mg²⁺, 0.15 mmol·L^-1 dNTPs, 0.60 μmol·L^-1 primers, 2.0 U Taq DNA polymerase and 26 ng template DNA could be able to amplified the most rich polymorphism and clear bands. This reaction system was experimentally validatied and and primers selected, and should be a suitable system for the genetic diversity analysis of I.indigotica Fort.

Ultrasound-assisted Extraction of Flavonoids from Calamus simplicfoilius Leaves and Its in vitro Antioxidant Activity

LIU Wei1,2；LI Yuan-Yuan1,2；WANG Hong-Zheng1,2*；YANG Lei1,2；ZU Yuan-Gang1,2

2015, 35(2):  315-320.  doi:10.7525/j.issn.1673-5102.2015.02.021

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We studied the total flavonoids extraction from leaves of Calamus simplicfoilius using ultrasonic-assisted extraction technology. A 3-factor, 3-level central composite design involving the three independent variables followed by response surface analysis was performed for the further optimization of the extraction parameters. The optimal extraction conditions were 72.74% of ethanol volume fraction, 22.5 mL·g^-1 of liquid-solid ratio, and 30 min of extraction time. Under these conditions, the experimental extraction of total flavonoids yield was 19.67 mg·g^-1. Ultrasonic-assisted extraction could reach more flavonoids and less time and energy comparing with traditionally extraction methods including macerate extraction and heat reflux extraction. By IR spectrums, the extracts contained amounts of flavonoids which had similar structure with lutein. Additionally, the DPPH free radical scavenging activity, reducing power and iron reduction activity of total flavonoids extracted using ultrasonic-assisted technology were a little lower than vitamin E, and the superoxide anions scavenging activity was significantly higher than vitamin E.