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    25 May 2019, Volume 40 Issue 5 Previous Issue    Next Issue
    Crop Culture and Nutrition, Genetic Breeding
    Nitrogen Nutrition Characteristics of Rubber Tree Leaves and Its Response to Nitrogen Application Rate
    CHEN Yanbin,ZHANG Yongfa,WANG Wenbin,XUE Xinxin,LUO Xuehua,WU Xiaoping,WANG Dapeng,ZHAO Chunmei
    2019, 40 (5):  831-838.  doi: 10.3969/j.issn.1000-2561.2019.05.001
    Abstract ( 962 )   HTML ( 47 )   PDF (813KB) ( 498 )  

    Rubber tree saplings planted after 2 years were used as the planting material, and a single-factor randomized block design field experiment was conducted to set three treatments with different nitrogen levels (low nitrogen 28 g/plant, medium nitrogen 56 g/plant and high nitrogen 84 g/plant). The parameters of rainfall, daily average temperature, nitrogen content of mature leaves, chlorophyll content and other indicators were dynamically observed for regression analysis and correlation analysis. The nitrogen content of rubber tree leaves showed a decline trend during the annual growth cycle. The difference of nitrogen contents in the leaves between different treatments was not significant. The dynamic change of chlorophyll content in the leaves had obvious seasonal variations, and several peaks were observed throughout the growth cycle. There was no significant chlorophyll content difference between different nitrogen levels. The results of correlation analysis showed that chlorophyll content and total nitrogen content were positively correlated (P<0.05). The nitrogen contents in the leaves was positively correlated with rainfall and temperature, and its correlation with temperature was extremely significant (P<0.01) while its correlation with rainfall was significant (P<0.05). Chlorophyll a, chlorophyll b and chlorophyll content were all extremely correlated with rainfall and temperature (P<0.01). Chlorophyll a/b ratio was extremely negatively correlated with N application rate, and a decrease in chlorophyll a/b ratio was observed with an increase in N rate. Obvious increment of N content in rubber leaves were found after N fertilizer application. Chlorophyll content could be regarded as an indicator revealing N status of rubber leaves to some degree. However, season and climate factors should be considered together.

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    Effects of Sisal Stalks Returned to the Field with Nitrogen Fertilizer on the Soil Fertility and Sisal Growth
    TAN Shibei,XI Jingen,ZHENG Jinlong,HE Chunping,WU Weihuai,LIANG Yanqiong,HUANG Xing,LI Rui,YI Kexian
    2019, 40 (5):  839-849.  doi: 10.3969/j.issn.1000-2561.2019.05.002
    Abstract ( 569 )   HTML ( 7 )   PDF (498KB) ( 305 )  

    In order to return the sisal stalks properly as the organic fertilizer, a pot experiment was carried out to study on the effects of sisal stalks returned to soil with nitrogen fertilizer application on the soil fertility and sisal growth. The results showed that the soil organic matter content, alkali hydrolysable nitrogen content and the soil urease activity were significantly higher than that in the controls, increased by 180.0%, 46.8% and 109.7%, respectively. And the three indicators also increased significantly when using the sisal stalks combined with different amount of urea. Soil invertase activity was significantly higher thanthat in the controls by using the sisal stalks combined with 1/2 amount of urea. Sisal leaf length after sisal stalks returned to the soil with 1/4 amount of urea was significantly higher than that in the control, the single application of urea and the single application of sisal stalks by 60.9%, 25.0% and 22.5%, respectively. Sisal stalks could replace some urea, and it needn’t use the full dose of urea when sisal stalks were returned to the field.

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    Evaluation on the Capacity of Different Organic Phosphorus Utilization in Reyan No. 4 King Grass
    HUANG Rui,ZHAO Xingkun,YU Daogeng,LIU Pandao,LIU Guodao,WANG Wenqiang
    2019, 40 (5):  850-856.  doi: 10.3969/j.issn.1000-2561.2019.05.003
    Abstract ( 656 )   HTML ( 11 )   PDF (1108KB) ( 316 )  

    In this study, Pennisetum purpureum × P. glaucum cv. Reyan No. 4 (King grass) was used to investigate the capacity to utilize four types of organic phosphorus (P) in a sand cultivation experiment. The results showed that King grass had different capabilities to mobilize adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), glucose-6-phosphate (G6P), and more capable of utilizing phosphate monoester (AMP or G6P) as the sole P source. Analysis of acid phosphatase (ACP) activity showed that total internal-ACP and secreted-ACP activities of roots significantly increased by more than 0.9-fold in organic P treatments. However, correlation between root total ACP activity and capacity to utilize organic P in King grass was not significant. Furthermore, significant positive correlation was found between root specific secreted-ACP activity and plant dry weight in different organic P treatments, with the observed R 2=0.861. Taken together, our results suggest that root specific secreted-ACP activity control organic P utilization in King grass. Our results would provide a theoretical basis for selecting and breeding tropical grass varieties with high P efficiency.

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    Effects of Different Medium and Trace Elements on Areca catechu L. Results and Yield Characteristics
    WANG Fengtang,YANG Fusun,CHEN Caizhi,LI Peizheng,BU Xianpan,JIU Fengfeng
    2019, 40 (5):  857-863.  doi: 10.3969/j.issn.1000-2561.2019.05.004
    Abstract ( 811 )   HTML ( 46 )   PDF (519KB) ( 481 )  

    In order to improve the yield and quality of areca (Areca catechu L.), we explored the effect of trace elements on the fruit production of areca. Seven medium and trace element fertilizers were applied to areca at the flowering and fruiting stages to determine the fruit setting rate, fruit shape and fruit quality. Trace elements significantly promoted the fruit set rate of areca. Magnesium significantly increased fruit set rate in small fruit and green fruit harvesting period, and promoted the yield per plant. Zinc, copper increased single fruit weight and single shuttle fruit yield significantly. Copper increased yield per plant by 41.23% significantly. Medium and trace element fertilizers improved the quality of areca. The fruit type ratio was 0.8951, 0.8139, and the fruit was full and round after iron and manganese use. Calcium increased the cellulose content in by 47.28% in the fruits. Iron promoted the vitamin C content. Magnesium significantly increased the content of soluble sugar of the fruit. Principal component analysis obtained the scores of five main indicators affecting the yield and quality. The effect of zinc fertilizer on the yield and quality was extremely significant. The comprehensive evaluation score was 1.8299 and 2.4893, respectively, followed by magnesium fertilizer. Copper fertilizer only increased the yield, and manganese fertilizer only improved the quality. It indicated that the treatment of medium and trace elements could increase the yield and quality of areca, but the effects of different elements were not the same.

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    Effects of Optimized Fertilization on Yield of Line Pepper and Utilization of Nitrogen in Latosols in Hainan
    JU Shengrong,WANG Wenbin,GENG Jianmei
    2019, 40 (5):  864-868.  doi: 10.3969/j.issn.1000-2561.2019.05.005
    Abstract ( 600 )   HTML ( 13 )   PDF (445KB) ( 315 )  

    Excessive application of nitrogen (N) fertilizer is very common to get high yield in vegetable production which causes water or air pollution. Compared with traditional N fertilizer application, the effects of reduced N fertilizer on the yield and accumulation of nitrogen in line pepper were studied by field experiments and the fate of N fertilizer was observed by the 15N micro-plot. The results showed that the line pepper yield by reduced 30% N was slightly improved but the difference was not significant compared with conventional N fertilizer input of 450 kg/hm 2. The results of the 15N micro-plot experiment indicated that the accumulation of nitrogen in fruits, stems and leaves was almost equal during the whole growth period of line pepper even though more N fertilizer was applied in conventional N fertilizer treatment which resulted in the increment of the residual N in 0-40 cm soil and the loss of N. The residual N may be leached after the harvest of line pepper and contributed to environment pollution. The nitrogen use efficiency of reduced 30% nitrogen was significantly higher than that of conventional N fertilizer obtained by the percentage of fertilizer N uptake by crop to N fertilizer rate or 15N tracer technique. Therefore, compared with conventional N fertilizer input of 450 kg/hm 2, it was necessary to reduce 30% nitrogen fertilizer in the line pepper production in Hainan, which would not affect the yield, and reduce environmental pollution.

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    Evaluation of Cold Tolerance of Mutant Offspring of Bermudagrass
    ZHANG Xiaoxiao,WANG Kai,GUO Hailin,WANG Rui,LI Dandan,LI Ling,LIU Jianxiu,GUO Aigui
    2019, 40 (5):  869-874.  doi: 10.3969/j.issn.1000-2561.2019.05.006
    Abstract ( 537 )   HTML ( 12 )   PDF (628KB) ( 376 )  

    According to morphological identification, 12 mutant progenies of ‘Yangjiang’ Bermudagrass with high turf value and low inflorescence density were selected to identify the cold tolerance by electrolyte leakage rate and stolon regrowth experiments. The electrolyte leakage rate showed that the variation range of LT50 ranged from -7.6 ℃ to -0.2 ℃, and the difference between the minimum and maximum values was 7.4 ℃. It was obvious that the cold resistance of the mutant offspring varied greatly. The order of cold resistance from strong to weak was M18>M4>M26> M28 >M22>Yangjiang>M29>M31>M10>M37>M16>M1>M25. Five of the mutants had better cold resistance than the parents, namely M18, M4, M26, M28 and M22. The results of stolon recovery test showed that the mutant growth rates of M1, M22, M26, M31 and M25 were higher than those of the parents after low temperature stress at 0 ℃and -5 ℃, and the recovery ability was better than that of the parents. Under -5 ℃low temperature stress, the ability to restore growth of M10, M37 and M28 was lower than that of the parents, and the cold resistance was relatively weak. Under low temperature stress of 0 ℃ and -5 ℃, the growth rates of M16 and M4 were lower than that of the parents, and the cold resistance was significantly weaker than that of the parents.The results of the two methods showed that the mutants M1 and M25 had strong recovery ability. M4 and M28 had better cold resistance and longer green period. M22, M18 and M26 had stronger cold resistance and stolon recovery ability. The cold resistance of the leaves of M29, M31 and the recovery ability of the stolons were similar to those of the parents. The cold resistance and the recovery ability of the stems of M10, M16 and M37 were weak, and the overall cold resistance was weak.

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    Phytophysiology and Biochemistry
    Effects of Chilling on Photosynthesis and Photosynthetic Pigment Contents in Leaves of Different Sugarcane Varieties
    ZHU Pengjin,PANG Xinhua,TAN Qinliang,LIANG Chun,YAN Lin,ZHOU Quanguang,CHENG Qin,LYU Ping,OU Kewei,LU Yefei,NING Lei
    2019, 40 (5):  875-881.  doi: 10.3969/j.issn.1000-2561.2019.05.007
    Abstract ( 577 )   HTML ( 14 )   PDF (776KB) ( 454 )  

    Using four sugarcane varieties (lines) with different cold resistance as the materials, the research analyzed the content and composition of the photosynthetic pigments in the leaves and used the chlorophyll fluorescence imaging to explore the influence of low temperature on the photosynthetic physiological characteristics in sugarcane. The content of chlorophyll of GR2 and GT28 per unit area was 13.74% and 10.39% higher than that of ROC22 respectively. The Fv'/Fm' was 3.25% and 3.01% higher, respectively, and the difference gradually increased with the prolongation of stress time, and ΦpsII and qP also had the same trends. The carotenoid content per unit area was 22.73% and 19.36% higher, NPQ was 20.33% and 12.17% higher in GR2 and GT28 than that of ROC22, respectively, and NPQ also increased rapidly with the extension of stress. The photosynthetic pigment content per unit leaf area of sugarcane at the early response stage (0-24 h) and the content of photosynthetic pigment per unit leaf area was basically unchanged, but Fv'/Fm', ΦpsII and qP in the leaf were inhibited, showing a rapid decrease and then relatively stable, while NPQ increased rapidly. At the adaptation stage (24-168 h), the leaf photosynthetic pigments were structurally destroyed due to low temperature stress, and Fv'/Fm', ΦpsII and qP were significantly reduced during 24-72 h since the treatment. However, at the treatment stage (72-168 h), the absorption, utilization and dissipation fractions of light energy were relatively stable, while NPQ increased sharply and then gradually became stabilized. The results indicated that under low temperature stress, sugarcane regulated the content of photosynthetic pigment and the combination ratio of PSI and PSII reaction center complexes to accelerate the dissipation of excessive excitation energy, adjusting the absorption of light energy by the photosynthetic apparatus, which was the adjustment on the absorption and utilization of light energy made by the photosynthetic apparatus under low temperature stress conditions and it was considered to be an important protection strategy of plants.

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    Effects of NaCl Stress on Some Physiological and Biochemical Indices and Photosynthetic Physiology Characteristics of Rice Cultivars with Different Salt Tolerance
    WANG Xuming,ZHAO Xiaxia,ZHOU Hongkai,CHEN Jingyang,MO Junjie,XIE Ping,YE Changhui
    2019, 40 (5):  882-890.  doi: 10.3969/j.issn.1000-2561.2019.05.008
    Abstract ( 574 )   HTML ( 13 )   PDF (653KB) ( 466 )  

    Two salt tolerances rice varieties ‘HH11’, ‘JX99’ and a salt-sensitive rice variety‘YSXD’ were cultured by a pot experiment until booting stage under soil salinity stress of 0, 1, 2, 3, 4 g/kg in a rainproof greenhouse. The study aimed to analyze the effects of NaCl stress on the physiological and biochemical indices and photosynthetic characteristics of rice varieties under salinity stress. Salinity stress inhibited the growth of rice, and the plant height of rice decreased with the increase of NaCl concentration, but the plant height of salt-tolerant rice was significantly higher than that of the salt-sensitive one under soil salt treatments of 3-4 g/kg. The response of soluble total sugars of the two salt-tolerant rice varieties and salt-sensitive rice variety to NaCl stress showed significant differences. The soluble total sugar of the salt-tolerant rice varieties was significantly higher than that of the salt-sensitive rice variety, but that of salt-sensitive rice variety significantly decreased under 1-4 g/kg NaCl concentration. The content of malondialdehyde in the leaves increased with an increase of NaCl concentration, which resulted in increased cell membrane permeability. The content of malondialdehyde in salt-tolerant rice varieties was less than that in salt-sensitive rice variety, and the cell membrane permeability of the salt-tolerant rice varieties was significantly lower than salt-sensitive rice variety. NaCl stress inhibited the photosynthetic rate of rice,but the reasons for the drop of photosynthetic rate caused by different NaCl concentrations were different. The decline of photosynthetic rate in salt-sensitive rice varieties resulted from non-stomatal limitation under 0-1 g/kg NaCl concentration, but which was caused by stomatal limitation under 2-4 g/kg NaCl concentration. The decline of photosynthetic rate in salt-tolerant rice variety HH11 resulted from non-stomatal limitation under 0-2 g/kg NaCl concentration, but which was caused by stomatal limitation under 3-4 g/kg NaCl concentration. Stomatal limitation was the main reason for the drop of photosynthetic rate of salt-tolerant rice variety JX99 under 0-2 g/kg NaCl concentration. Leaf transpiration rate in salt-tolerant rice variety was significantly decreased, and that was significantly lower than that of the salt-sensitive rice variety, but water use efficiency and stomatal limited value were significantly increased and were significantly higher than that of the salt-sensitive rice varieties under salinity stress. The results showed that salt concentration increased stomatal limited value, reduced water loss, inhibited transpiration rate and improved leaf water use efficiency to meet the needs of normal metabolic physiology of salt-tolerant rice varieties to maintain normal life activities.

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    Effects of Low Temperature Stress on the Photosynthetic Characteristics and Chlorophyll Fluorescence Parameters of Bletilla striata
    CUI Bo,CHENG Shaoli,YUAN Xiuyun,ZHOU Yiran,HAO Ping’an,LI Junlin,MA Jie
    2019, 40 (5):  891-897.  doi: 10.3969/j.issn.1000-2561.2019.05.009
    Abstract ( 555 )   HTML ( 15 )   PDF (474KB) ( 370 )  

    In order to study the physiological responses to low temperature stress in Bletilla striata, one-year-old B Striata plants were treated by four different low temperatures of 22, 18, 14 and 10 ℃, respectively. The low temperature treatment was conducted during the subsequent 0, 7, 14 and 28 days, whereas the plant recovery treated for 3 and 7 days. Samples from the all above treatments were used to measure the photosynthetic characteristics and chlorophyll fluorescence parameters. The net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) of the leaves decreased significantly under 14 and 10 ℃ treatment while the intercellular carbon dioxide concentration (Ci) decreased at first and then increased with the lowest value at 14 d. The chlorophyll fluorescence parameter Fo increased and the other three indexes including Fv/Fm, Y(II), qP and ETR decreased significantly, respectively. These findings indicted that the PSⅡ photosynthetic system of B. Striata was damaged by 10 and 14 ℃ temperature treatments, and further to affect the plant growth. The Pn, Gs, Tr and Ci values decreased slowly under 18 and 22 ℃ treatments, indicating that the decrease of Pn value was leaded by the stomatal factors. Moreover the index Fo increased slightly but Fv/Fm, Y(II), qP and ETR decreased slightly, suggesting that the PSⅡ photosynthetic system was damaged slightly. Finally, the changes of Y(II), qP, ETR, Fo, Fv/Fm were not obvious under the 22 ℃ treatment, indicating the photosynthetic system was not damaged by the low temperature treatment. These findings suggested that the growth and photosynthesis of B. striata were severely inhibited by 10 ℃ and 14 ℃ low temperature treatments, and the plant morphology was not affected by the 18 ℃ treatment, but the photosynthesis was inhibited.

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    Biotechnology and Tissue Culture
    Cloning of HbHsfA4a and Its Transcriptional Responses to Cold Stress in Hevea brasiliensis
    YU Wencai,LI Yan,WU Shaohua,CHAO Jinquan,ZHANG Shixin,YANG Shuguang,TIAN Weimin
    2019, 40 (5):  898-904.  doi: 10.3969/j.issn.1000-2561.2019.05.010
    Abstract ( 558 )   HTML ( 15 )   PDF (4855KB) ( 474 )  

    Heat shock transcription factors (Hsfs) play an important role in the response of plants to abiotic stresses. An Hsf gene, designated as HbHsfA4a, was cloned from a rubber tree (Hevea brasiliensis) clonal line 93-114 by RT-PCR. The ORF of HbHsfA4a was 1215 bp in length which would encode 404 amino acid residues. The putative protein was predicted to possess a molecular mass of 46.40 ku and an isoelectric point of 4.91. Quantitative Real-time PCR (QRT-PCR) analysis showed that the expression of HbHsfA4a was highly induced in the bark tissue of 93-114 by cold stress (4 ℃) and significantly higher than in Reken501 under the same cold treatment. Further analysis showed that the level of the background expression and the expression of HbHsfA4a in response to cold stress within 8 h in the bark tissue of the two cold-tolerant rubber tree clones was significantly higher than that of the two cold-sensitive rubber tree clones. The up-regulation of HbHsfA4a might be crucial to the survival of rubber trees under cold stress.

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    Cloning and Functional Characterization for Abiotic Stress Tolerance of TtASR Gene from Tetragonia tetragonioides
    YE Yuyan,LUO Ming,CHEN Hongfeng,ZHANG Mei,HE Jinshi,YANG Lixiang
    2019, 40 (5):  905-912.  doi: 10.3969/j.issn.1000-2561.2019.05.011
    Abstract ( 865 )   HTML ( 12 )   PDF (3530KB) ( 403 )  

    ABA, stress and ripening (ASR) protein belongs to a plant-specific small family of proteins with low molecular weight and high hydrophobicity, and is supposed to play protective roles in plant cells under abiotic stresses. An ASR gene, designated as TtASR (GenBank accession no.: MH454101), was isolated from a cDNA library prepared from Tetragonia tetragonioides (Pall.) Kuntze and both the nucleotide and the deduced protein sequences were analyzed. The full length cDNA of TtASR contained a 723 bp long open reading frame, which would encode a peptide of 240 amino acid residues with a molecular weight of 25.29 ku and an isoelectric point of 5.26. TtASR shared a high amino acid sequence homology with the ASRs from Suaeda liaotungensis (SlASR) and Salicornia brachiata (SbASR-1). The open reading frame of TtASR was inserted into the prokaryotic expression vector of pGEX 6p-1 and transformed into E. coli. The E. coli cells under the induction of IPTG showed higher tolerance to NaCl, osmotic stress and oxidative stress than control cells.

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    Cloning and Functional Analysis of NtNAC1 Gene in Narcissus tazetta ‘Yunxiang’
    XIAO Yaoyu,LI Lin,LI Quanchao,LIU Yang,CHEN Xiaojing
    2019, 40 (5):  913-921.  doi: 10.3969/j.issn.1000-2561.2019.05.012
    Abstract ( 523 )   HTML ( 15 )   PDF (15185KB) ( 135 )  

    NtNAC1 was cloned by RT-PCR from the roots of Narcissus tazetta ‘Yunxiang’. The gene was further characterized by analyzing its tempo-spacial expression patterns in N. tazetta ‘Yunxiang’ plants, as well as changes in its expression in the leaves and roots under different stresses and stress-related hormones, and by analyzing the NtNAC1 over-expressing transgenic tobacco plants. The results showed that the cloned full-length cDNA was 1083 bp long, containing an ORF of 918 bp long which would putatively encode a peptide of 306 amino acid residues. The deduced protein possessed a highly conserved DNA binding domain, NAM, at its N-terminal, and was grouped, together with its most closely related proteins, MusaSNAC1 and TaNAC47, into the SNAC1 subgroup on the phylogenetic tree. Quantitative real-time PCR results showed that the expression level of NtNAC1 in petals and coronas first increased and then decreased as the floral organs developed, and reached a peak in the early flowering period. NtNAC1 was abundantly expressed in roots and leaves, and was induced by the treatments of ABA, MeJA, SA, H2O2, NaCl and PEG, indicating that it may be involved in the morphogenesis of floral organs in addition to its principal role in the responses of plants to abiotic stresses. Nine transgenic tobacco lines carrying NtNAC1 were obtained by agrobacterium-mediated transformation and three of them were over-expressing lines. The NtNAC1 over-expressing lines showed better developed root systems and higher survival rates under the treatments of high concentrations of salt and mannitol, as compared with the non-transgenic plants. Our results showed that expressing NtNAC1 increased stress tolerance of plants and its potential in the improvement of abiotic stress resistance of N. tazetta could be further exploited.

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    Cloning of Ascorbate Peroxidase Gene and Analysis of Its Gene Expression and Enzyme Activity of Somatic Embryogenesis in Camellia sinensis
    GUO Yuqiong,HUANG Daobin,LI Xiaozhen,ZHU Chen,ZHANG Shuting,FU Haifeng,ZHOU Chengzhe,OUYANG Mingqiu,CHEN Lan,LAI Zhongxiong
    2019, 40 (5):  922-931.  doi: 10.3969/j.issn.1000-2561.2019.05.013
    Abstract ( 551 )   HTML ( 13 )   PDF (4892KB) ( 277 )  

    The ascorbate peroxidase gene CsAPX (GenBank accession number: MG799534.1) was cloned from the somatic embryo of Camellia sinensis cultivar ‘Tieguanyin’. The CsAPX gene contained an open reading frame (ORF) of 753 bp in length, which would encode a total of 250 amino acids, and bioinformatics analysis about CsAPX gene was also employed. The total APX activity was measured during the whole somatic embryogenesis and the results showed that the total APX enzyme activity presented a downward trend in the globular embryo to cotyledonary embryo stage, and reached the lowest value in the cotyledon embryo stage. With the further differentiation of the embryo cells, the total APX enzyme activity increased continuously during the cotyledonary embryo stage, the embryo maturation stage and the germinating stage, and showed an overall trend of “decreasing first and then rising”. The expression of CsAPX was significantly different between the 5 stages of the somatic embryogenesis, showing an overall trend of “up-down- up” with a significantly higher expression level in the germinating stage than in the other 4 embryo stages. It was speculated that CsAPX played an important role in the germinating stage.

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    Crop Pests, Diseases and Their Control, Environmental Protection
    Cloning and Prokaryotic Expression of CsHog1 Gene from Colletotrichum siamense
    LIAO Xiaomiao,HE Qiguang,LIU Yao,XU Liangxiang,ZHOU Xiaoyun,LIU Wenbo,ZHANG Yu,LIN Chunhua,MIAO Weiguo
    2019, 40 (5):  932-938.  doi: 10.3969/j.issn.1000-2561.2019.05.014
    Abstract ( 591 )   HTML ( 9 )   PDF (4186KB) ( 307 )  

    The high osmolarity glycerol (HOG) response pathway plays important roles in growth and development, pathogenesis, fungicide sensitivity etc. in phytopathogenic fungi, but its function may vary in different pathogenic fungi. In this study, sHog1 was cloned by homologous cloning from Colletotrichum siamense isolated from rubber trees in order to understand the role and regulatory mechanisms of Hog1 protein. GST-Hog1 fusion protein expression vectors were also constructed and expressed in Escherichia coli BL21 (DE3). The expressed fusion proteins were analyzed on SDS-PAGE and Western Blotting. The results showed that CsHog1 contained 5 introns and encoded 459 amino acids and had the Serine / Threonine protein kinases catalytic domain S_TKc. Phylogenetic clustering showed that CsHog1 shared the highest similarity to ClOsc1 (Hog1 homologue) of C. lagenarium. SDS-PAGE analysis showed that GST-Hog1 protein was successfully expressed in E. coli under the induction of IPTG at 0.3, 0.5, 0.8 and 1.0 mmol/L and at incubation temperature of 16, 25 or 28 ℃. The size of the fusion proteins was 70 ku which was identical to the expected molecular weight. Western Blotting showed that the purified fusion protein could be recognized by GST antibodies.

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    Proteomic Analysis of Different Sugarcane Genotypes in Response to Pokkah Boeng Disease
    WANG Zeping,LIN Shanhai,LIANG Qiang,LI Changning,SONG Xiupeng,LIU Lu,LI Yijie
    2019, 40 (5):  939-946.  doi: 10.3969/j.issn.1000-2561.2019.05.015
    Abstract ( 469 )   HTML ( 13 )   PDF (677KB) ( 394 )  

    This study was to provide a theoretical guidance and scientific reference for sugarcane resistance breeding and disease prevention by analyzing the physiological and biochemical mechanism in the infection process of sugarcane response to pokkah boeng disease. With spore suspension of Fusarium verticillioides (the mainly pathogenic fungus of pokkah boeng disease in China) as the pathogeny and YT94/128 (HR), GT37 (HS) as the tested materials by syringe inoculation under a greenhouse, and the samples were collected at the 14 th post-inoculation with the highest disease index to carry out quantitative expression analysis by isobaric tags for relative and absolute quantitation (iTRAQ). The results showed a total of 3707 proteins were successfully identified from GT37, and 542 differentially expressed proteins (DEPs) were obtained, of which 187 were up-regulated and 355 were down-regulated. Meanwhile, a total of 3068 proteins were successfully identified from YT94/128, and 449 DEPs were obtained, of which 191 were up-regulated and 258 were down-regulated. In a word, there are significant differences in the protein composition of sugarcane varieties with different genetic backgrounds, and it is speculated that this is the important molecular basis for the difference of resistance on pokkah boeng disease between different sugarcane varieties.

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    Scanning Electron Microscope Observation on the Changes of Litchi Exocarp Inoculated with Phytophthora litchii
    SUN Jinhua,CAO Lulu,LI Huanling,WANG Guo,WANG Shujun,LI Fang,WANG Jiabao,ZHANG Lei
    2019, 40 (5):  947-952.  doi: 10.3969/j.issn.1000-2561.2019.05.016
    Abstract ( 623 )   HTML ( 16 )   PDF (5774KB) ( 433 )  

    Litchi downy blight is a damaging oomycete disease, caused by Phytophthora litchii. The 4×10 4cfu/mL sporangium suspension was used to inoculate the fruit of the resistant cultivar ‘Heiye’ and susceptible cultivar ‘Guiwei’. The ultra-structural changes of exocarps was investigated by the Scanning Electron Microscopy (SEM) method at 0, 1, 6, 12, 24, 48, 72, and 96 h post-inoculation. The results indicated that the lobe of exocarp was flat and consisted of convex and compact honeycomb in ‘Heiye’, while the valley of lobe was deep, and the honeycomb was thin and erect in ‘Guiwei’. The integrity of exocarps was damaged with the prolonging of the harvest time. The infection of P. litchii promoted the dissociation of exocarps, as well as the damage degree of ‘Hieye” was lower than that of ‘Guiwei’. Under the same inoculation condition, the exocarps of ‘Guiwei’ harbored nearly five times sporangia than that of ‘Heiye’. The character of lobe and honeycomb was more easily agglutinated and infected by the spores of P. litchii in ‘Guiwei’. The infection of P. litchii accelerated the breakdown of exocarp structure, which had a small impact on ‘Heiye’. Comparing with ‘Guiwei’, the gemination of spores, as well as growth of mycelium and formation of sporangia, was lagging in ‘Heiye’. It is suggested that the growth of P. litchii is repressed in the exocarps of resistant cultivars. The structure difference of the exocarp is the reason why ‘Guiwei’ is more susceptible to P. litchii than ‘Heiye’.

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    Species and Parasitic Characteristics of Parasitic Wasp of Mulberry Pyralid Glyphodes pyloalis Walker (Lepidoptera:Pyralidae) in Hainan
    WANG Nayu,LU Fuping,GENG Tao,WU Huazhou,WANG Yongsong,ZHU Jiawen,WANG Shuchang
    2019, 40 (5):  953-959.  doi: 10.3969/j.issn.1000-2561.2019.05.017
    Abstract ( 475 )   HTML ( 7 )   PDF (442KB) ( 369 )  

    It is very important to find out the natural enemy species and the biological characteristics of Glyphodes pyloalis Walker, one of the main mulberry pests, for improving efficiency of biological control. The investigations on the parasitic wasps of G. pyloalis in Hainan, China was carried out. Five parasitic wasps of G. pyloalis were described. They were Apanteles heterusiae Wilkinson, Phanerotoma planifrons Nees, Chelonus pectinophorae Cushman, Macrocentrus philippinensis Ashmead and Brachymeria lasus Walker. Among them, the dominant species was Apanteles heterusiae, which had the highest parasitic rate of 63.24%. A. heterusiae was gregarious parasitoids and the numbers of its offspring per female was 3-16. The other four parasitic wasps were all solitary parasitoids and the average parasitic rate in the field was 9.15%, 6.21%, 5.62% and 5.24%, respectively. A. heterusiae mainly parasitized the 1 st and 2 nd larvae of G. pyloalis and its parasitic rates get a higher level when the individual number of the 1 st and 2 nd larvae of G. pyloalis was 9 and 8, respectively. Occasionally the 3 rd larvae of G. pyloalis was also been observed being parasitized. Mature larvae gnawed out of body wall of the 5 th larvae of the host and pupated in 5-7 hours completely. Then the adults emerged with an emergence rate 33.33%-100% after 4-5 d. The sex ratio was 0-87.50% and the life span always 1 to 3 days and that of the male was slightly shorter than that of the female. In the similar experiment replaced G. pyloalis with the silkworm, no parasitism was observed. This study would provide precondition for the artificial reproduction of A. heterusiae.

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    Population Reproductive Characteristics of Opisina arenosella Walker
    LIN Yuying,JIN Tao,MA Guangchang,WEN Haibo,JIN Qi’an,PENG Zhengqiang
    2019, 40 (5):  960-964.  doi: 10.3969/j.issn.1000-2561.2019.05.018
    Abstract ( 543 )   HTML ( 11 )   PDF (574KB) ( 331 )  

    The population reproductive characteristics of Opisina arenosella Walker were investigated. O. arenosella was collected from the wild field of Danzhou, Hainan every month in 2015. The female and male adults on the day when emerging and the preoviposition duration, egg number per female, hatching rate of eggs, newly hatched larvae and longevity of adults were recorded. The temperature was the lowest in January, and then the preoviposition duration which was (3.00±0.21)d in January was the longest. The preoviposition durations in May and June took the second place as the temperatures were higher, and the preoviposition durations in other months were shorter. The egg number per female which was (231.07±19.55) eggs in November was the highest, the egg number per female in March, April, July, August, September, October was lower, and the egg number per female was the lowest in January, February, May, June, December. The hatching rate of eggs which was (2.70±2.01)% in January (low temperature) was the lowest, and the difference in hatching rates of eggs from February to December was not significant. The number of newly hatched larvae which was (4.17±2.94) individuals in January was the least, less in May, June which were (54.04±8.18) and (52.31±10.81) individuals, respectively and the most in March which was (87.89±14.94) individuals. The longevity of adults was extended in low temperature, the longevity of female adults of each month was shorter than that of male adults, and the average life longevity of male adults in May and June which was (7.08±0.68) and (7.67±0.84)d, respectively and female adults in May and June which was (6.50±0.34) and (6.22±0.36) d, correspondingly was shorter than that in other months. High temperature in May and June would inhibit the population of O. arenosella, so it was the best time for chemical control of the insect. This study would provide a basic scientific basis for the prevention and control, and the field forecast of O. arenosella.

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    Agricultural Product Processing, Preservation, Storage, Analysis and Detection
    Analysis of Volatile Compounds in Guanyin Tea Stem
    WANG Wei,HUANG Xujian,LIN Yunzhi,TANG Rongjin,GUO Yaling
    2019, 40 (5):  965-972.  doi: 10.3969/j.issn.1000-2561.2019.05.019
    Abstract ( 486 )   HTML ( 12 )   PDF (452KB) ( 358 )  

    The tea stem of Anxi Guanyin was taken as the research object and the volatile substances of stem were extracted and analyzed by headspace sampling-gas chromatography/mass spectrometry (HS-GC/MS). Its qualitative analysis was achieved by mass Spectrometry (MS) combined with retention index (RI). The type and relative content of the volatiles and the biological activity of some volatiles were compared and discussed. 130 compounds were detected in the tea stem of Anxi Guanyin, of which 108 were in common. 45 compounds were totally identified and 58 compounds were preliminarily identified by MS, while 54 compounds were identified by RI. 68 compounds were totally identified by MS combined with RI, meaning the combination of MS and RI greatly improved the quantity of qualitative volatiles and accuracy of quality. The types and contents of hydrocarbons, aldehydes, alcohols and esters were the most abundant in the stem. There were 24 volatiles with relative content higher than 1%. Some volatiles, whose content accounted for 20.51% of the total volatile content of tea stem, had biological activities, such as anaesthesia, bacteriostasis, anticancer and et al. The study would provide a theoretical basis for the role of the tea stem in the formation of aroma quality of Oolong tea and the comprehensive utilization of tea stems.

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    Effect of Physical Modification Treatment on Physico-chemical Properties of Modified Dietary Fiber from Pineapple Pomace
    JIANG Yongchao,LIN Lijing,GONG Xiao,HUANG Xiaobing,ZENG Yong,LI Jihua
    2019, 40 (5):  973-979.  doi: 10.3969/j.issn.1000-2561.2019.05.020
    Abstract ( 572 )   HTML ( 10 )   PDF (1109KB) ( 389 )  

    The dietary fiber from pineapple pomace was modificated by three physical modification methods including ultrafine grinding, steam explosion, and extrusion. Changes in the composition, physical, topographical structure, bioactive content, and other chemical properties of pineapple pomace dietary fiber were investigated. The three physical modification methods could improve the cation exchange capacity of the samples and increase the dissolution of the polyphenols and flavonoids. Among them, the steam explosion treatment could significantly increase the content of soluble dietary fiber, and it performed well in water and oil holding. The results of morphological structure analysis showed that the structure of the modificated dietary fiber changed in different degrees, but the major components and chemical structure were not affected. In general, the steam explosion treatment improved the quality of pineapple pomace dietary.

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    Preparation and Physicochemical Properties of Amphiphilic Chitosan/ Quercetin nanomicelles
    WANG Hui,YANG Ziming,HE Zuyu,ZHOU Chuang,WANG Chao,LI Puwang
    2019, 40 (5):  980-986.  doi: 10.3969/j.issn.1000-2561.2019.05.021
    Abstract ( 586 )   HTML ( 8 )   PDF (1196KB) ( 619 )  

    Quercetin (QCT) is a flavonoid compound with strong antioxidant and anti-inflammatory effects, which is widely found in tropical plants. However, the low solubility and poor stability of quercetin limit its application in food and medicine. In this study, a novel amphiphilic chitosan (ACS) with deoxycholic acid (DA) as the hydrophobic group and n-acetyl-l-cysteine (NAC) as the hydrophilic group was synthesized, and the amphiphilic chitosan quercetin nanoparticle micelle (ACS-QNMs) was prepared and characterized. FTIR showed that DA and NAC were grafted onto chitosan successfully. The critical micelle concentration (CMC) of blank micelles was 26.92 μg/mL. Scanning electron microscopy (SEM) was used to observe the spherical appearance and uniform distribution of nanoparticles. The micelle potential was determined by nano particle size potentiometer to be 31 mV. The results showed that quercetin could be released in vivo and stored stably at room temperature after being embedded in nanomicelles.

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    Optimization of Fermentation Technology for Yellow Wine from Acerola Cherry and Rice by Response Surface Methodology
    JU Xueli,TAN Haisheng,YANG Jinsong,GUO Haiyang,SHI Yike,WU Yunxue
    2019, 40 (5):  987-994.  doi: 10.3969/j.issn.1000-2561.2019.05.022
    Abstract ( 515 )   HTML ( 8 )   PDF (1592KB) ( 353 )  

    The Box-Behnken response surface method was used to optimize the fermentation process of yellow wine from acerola cherry and rice on the basis of single factor experiments. The best fermentation conditions were as follows: koji addition 0.8%, saccharification temperature 31 ℃, saccharification time 68 h, Saccharomyces cerevisiae 0.8%, liquid to material ratio 1.1: 1(mL: g), acerola cherry 33%, fermentation temperature 26 ℃, and fermentation time 4 d. Under the best fermentation conditions, the yellow wine produced had orange color, clear and transparent characteristics, harmonious and fragrant aroma, soft and mellow taste, alcohol content 13.8% vol, and the sensory evaluation score was 96.4. Fruit and grain blending is the development direction of the wine making for tropical fruits. The production of yellow wine from acerola cherry and rice would be of importance both in the development of new varieties of fruit wine and marketing.

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    Optimization of Liquid Fermentation Conditions of Bacillus subtilis CS27
    ZHANG Hui,LIN Chenqiang,WU Dahua,CHEN Jichen,CAI Haisong
    2019, 40 (5):  995-1001.  doi: 10.3969/j.issn.1000-2561.2019.05.023
    Abstract ( 598 )   HTML ( 10 )   PDF (536KB) ( 436 )  

    The fermentation medium and culture conditions were optimized by single factor and orthogonal experiments. The optimum fermentation medium was 1.5% soybean powder, 0.5% molasses, 0.8% ammonium chloride, 1.0% NaCl, 0.1% sodium citrate, 0.5% calcium carbonate, 0.1% MgSO4·7H2O. The optimum fermentation conditions were fermentation temperature 32 ℃, liquid volume 50 mL / 250 mL, inoculation amount 10%, initial pH value 7.0. The cell biomass could reach 1.75 × 10 9cfu/mL under the optimized fermentation medium, which could provide a scientific basis for the large scale production and application of B. subtilis CS27.

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    Screening of Insecticidal Activities of 16 Traditional Chinese Medicine Containing Lactam Derivatives of Aristolochia Against Musca domestica and Aedes aegypti
    YU Senquan,DONG Cunzhu,SU Lyu,GUO Jiaohong,HOU Zongmin
    2019, 40 (5):  1002-1007.  doi: 10.3969/j.issn.1000-2561.2019.05.024
    Abstract ( 490 )   HTML ( 7 )   PDF (504KB) ( 311 )  

    In order to find and develop safe and effective botanical insecticides, to enrich the sources of botanical pesticides and fill the gaps left by prohibitions and restrictions on the use of chemical pesticides, sixteen kinds of Chinese herbal medicines were extracted by methanol, and the insecticidal activity against adult Musca domestica was tested by the method of unlimited intake, and that against the fourth instar larvae of Aedes aegypti was tested by the larval immersion method. The results showed that after 48 h treatment at 10 mg/mL, the corrected mortality rates of Asarum heterotropoides F. Schmidt var. mandshuricum (Maxim.) Kitag, Piper wallichii (Miq.) Hand-Mazz, Aristolochia kaempferi Willd. [A.mollis Dunn], Asarum himalaicum Hook. f. et Thoms. ex Klotzsch was 100%, 100%, 100% and 98.33%, respectively, and the LC50 value separately was 0.1483, 0.1544, 3.5277 and 2.4292 mg/mL, while that for Derris trifoliata Lour was 0.9080 mg/mL. Against the 4 th instar larvae of A. aegypti at 0.5 mg/mL for 48 h, the corrected mortality rate of A. heterotropoides, P. wallichii, A. kaempferi, A. himalaicum, and Magnolia officinalis Rehd. et Wils. was 100%, 100%, 100%, 100% and 100%, separately, and the LC50 value was 0.0046, 0.0094, 0.0570, 0.0658 and 0.0504 mg/mL, respectively, while that for D. trifoliata Lour was 0.1194 mg/mL. A. heterotropoides, P. wallichii, A. kaempferi, A. himalaicum showed excellent insecticidal activity on the two tested insects, better than or close to the activity of the control D. trifoliata.

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    Ecology and Review
    Impacts of Reduced Fertilizer Application with Organic N on Vegetable Growth and Soil N Balance in Vegetable Fields in Latosolic Red Soil Zones
    NING Jianfeng,AI Shaoying,LI Mengjun,YAO Jianwu,YU Danni,WANG Siyuan,SHEN Jian
    2019, 40 (5):  1008-1014.  doi: 10.3969/j.issn.1000-2561.2019.05.025
    Abstract ( 496 )   HTML ( 12 )   PDF (553KB) ( 436 )  

    A plot experiment with six treatments (i.e. CK, unfertilized control; CF, conventional fertilization; OPT, optimized fertilization; ON, chemical fertilizer nitrogen substituted partially by organic nitrogen under OPT mode: T10, T20 and T30) was conducted in a perennial planting vegetable field in the latosolic red soil zone to investigate the effect of optimal fertilization with or without organic nitrogen substituting partial chemical fertilizer nitrogen on vegetable growth and soil nitrogen balance. Four successive crops with a leafy vegetable Pakchoi (Brassica?chinensis L.) were carried out. The results showed that the yields of Pakchoi increased significantly after different fertilizers applied, and the contribution rate of fertilizer on yield increase ranged from 23.1% to 39.6%. The yield of Pakchoi was 3169—3369 kg/667 m 2 and the plant total nitrogen uptake was 33.4 kg/667 m 2 with nitrogen use efficiency of 31.2% under CF treatment for four successive crop periods. As compared to CF, vegetable yields, plant nitrogen uptake and nitrogen use efficiency in treatment of OPT, T10, T20 and T30 showed no significant differences. It was estimated that chemical fertilizer application rate in OPT, T10, T20 and T30 was reduced by 35%, 38%, 41% and 44%, respectively. Nitrogen surplus in the Pakchoi planting system was 33.42 kg/667 m 2 under CF treatment, which decreased by 18%—48% with treatment of OPT, T10, T20 and T30. It was suggested that the reduction of 35%—44% in chemical fertilizer application rate under conventional fertilization mode was feasible based on the stability of vegetable production and lower soil nitrogen surplus and its potential risks of nonpoint source pollution. The partial chemical fertilizer nitrogen in OPT treatment substituted by organic nitrogen contributed to further reduction of fertilizer input, leading to more reduction of fertilization in the vegetable planting system. It was concluded that OPT or T10, T20 and T30 treatment should be considered as a recommendation for regional vegetable production in the latosolic red soil zone.

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    Heavy Metal Pollution and Potential Ecological Risk Assessment of Pogostemon cablin
    HUANG Weizhan,HU Zhenzhen,LU Changhua,ZHANG Hongyi,HE Mengling,YAN Hanjing
    2019, 40 (5):  1015-1021.  doi: 10.3969/j.issn.1000-2561.2019.05.026
    Abstract ( 481 )   HTML ( 7 )   PDF (339KB) ( 304 )  

    In this study, national standard methods were used to determine the concentrations of Cu, Pb, Cd, Cr, As and Hg in the medicinal materials of Pogostemon cablin and its habitat soil in 10 regions. The distribution of six heavy metals and the bioconcentration factors were analyzed by the Nonparametric Test. Then, the Single Factor Pollution Index (Pi), Nemerow Synthetical Pollution Index (Pcombined), and a Potential Ecological Risk Index (RI) were used to evaluate the pollution levels of the 10 soil samples. All habitat soils were qualified and the concentrations of heavy metals in P. cablin generally met the standard, while Hg in four producing areas exceeded the standard in varying degrees. Significant differences were obviously observed among the distributions of heavy metals Pb, Cu and Cd in different habitat soils, the concentrations of Pb, Cr and As in the medicinal materials and its habitat soil, as well as the bioconcentration factor of Pb in different areas of the medicinal materials. Pcombined and Pi showed that the soil in Sihui City, Cd in the soil in Changfeng County, Pb in the soil in Liantang Town, Pb and Cd in the soil in Sihui City were near the warning limit, while the other habitat soils were still clean. In terms of RI value, each production area were at slight pollution level. In addition, P. cablin from different habitats had an enrichment effect of Cd and Hg in the soil. Therefore soils with low level of heavy metals should be selected to ensure the safety of medicinal materials when farming P. cablin.

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    Investigation of Introduced Plants in Meiji Reefs, China
    HUANG Shengzhuo,DUAN Ruijun,WANG Jun,MEI Wenli,DAI Haofu
    2019, 40 (5):  1022-1031.  doi: 10.3969/j.issn.1000-2561.2019.05.027
    Abstract ( 825 )   HTML ( 7 )   PDF (496KB) ( 444 )  

    In order to know the condition of introduced plants in Meiji Reef, the southernmost of China, the introduced plants in the reef was investigated from July to Augest, 2018, and the species and growth status of these plants were analyzed with field investigation. There were 279 species (including varieties and forms) of introduced vascular plants, belonging to 220 genera and 75 families, more than these in 2012. Based on plants species in neighouring Xisha Islands, some plant species suitable for wind prevention and sand fixation, greening constructionin, and edible herbs, vigitables, and fruits in Nansha Islands (reefs) such as Meiji Reef were also proposed. It would provide references to greening construction, keeping ecological balance, providing vegetation and promoting military and civilian integration of Nansha Islands in the future.

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    Advances in the Regulation of Fruit Quality by AP2-ERF Transcription Factor
    SONG Kanghua,LI Yuting,ZHANG Lubin
    2019, 40 (5):  1032-1040.  doi: 10.3969/j.issn.1000-2561.2019.05.028
    Abstract ( 1004 )   HTML ( 74 )   PDF (421KB) ( 622 )  

    Ethylene is an important plant hormone that affects the quality of fruit. Studies of ethylene signal transduction pathway have been widely carried out in different kinds of fruits, but its transcriptional regulation mechanism is still at the beginning stage. This paper summarized the progresses in the transcriptional regulation of fruit quality based on the AP2/ERF results. Perspective and problems for further exploration of AP2/ERF on the fruit quality improvement were also discussed.

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