in vitro culture through investigating the influences factors which relate to phenolic browning and observing stem tip cellular compartments structure changes in the process of browning. Sugarcane variety ROC 22 which provided by sugarcane research institute of Guangxi Academy of Agricultural Sciences was selected as experiment material. The result showed that different axillary buds obtained obviously different induction survival rates, the more fresh buds resulted in the more high survival rate, the first position bud got the highest survival rate, that is to mean the more fresh bud get the more higher induction survival rate; the induction survival rate was not effected by different explants collecting seasons; pre-germination time 4 weeks resulted higher induction survival rate, and obtained significant difference compared with other pre-germination time treatments. Different buds with different pre-germination time treatments had obvious different total phenolic compounds content, the longer pre-germination time resulted in the lower phenolic compounds accumulated in stem tip cells, pre-germination time for 2 weeks and 3 weeks obtained higher total phenolic compounds in stem tip cells at different position buds, while pre-germination for 4 weeks and 5 weeks got low total phenolic compounds content, therefore, sugarcane axillary bud was pre-germinated for 4 weeks is a better period for induction tissue culture; stem tip total phenolic compounds content was not effected by explants collecting season; polyphenol oxidase acitivity changed significantly in different axillary buds with different pre-germination time treatments, lower polyphenol oxidase acitivity were obtained in 1-6 position buds with 2 weeks pre-germinated treatment and in 3-6 position buds with 4 weeks pre-germinated treatment; while higher polyphenol oxidase acitivity were obtained in 1-3 position buds with 3 weeks pre-germinated treatment and in 4-6 position buds with 5 weeks pre-germinated treatment. Sugarcane browning stem tips were observed by electron microscope, the result showed that the nucleus structure deformed, swollen mitochondria stretched, part of the vacuole membrane started to break down at the early browning stage; more serious plasmolysis appeared, a large number of lysosomes appeared in cytoplasm, organelles such as mitochondria completely decomposed, cell membrane, vacuole membrane, nuclear membrane and mitochondrial membrane broke down, cytoplasm completely dissolved, vacuoles mitochondria and other organelles completely decomposed at the later browning stage. This finding inferred that nucleus and mitochondria structural distortion and cell membrane systems break were the main reasons for sugarcane stem tip browning to death in the process of tissue culture in vitro.]]> 2016/5/31 17:34:54 2, TANG Yun-Xian¹, LIAO Fen¹, WANG Miao², LIANG Yong-Jian¹, YANG Li-Tao^1*, HUANG Dong-Liang², LI Yang-Rui^1,2]]> YANG Liu², TANG Yun-Xian¹, LIAO Fen¹, WANG Miao², LIANG Yong-Jian¹, YANG Li-Tao^1*, HUANG Dong-Liang², LI Yang-Rui^1,2 6true Lonicera macranthoides Hand.-Mazz. “Yulei 1”, this paper discussed that the different species of inorganic salt had many effects on the chlorogenic acid content and growth of cell in suspension cultures of L. macranthoides Hand.-Mazz. “Yulei 1”. We weighed the biomass of the L. macranthoides Hand.-Mazz. “Yulei 1” cells and analyzed the content of chlrogenic acid by HPLC through the additions of different concentrations of inorganic salt in cell suspension cultures of L. macranthoides Hand.-Mazz. “Yulei 1”. The result showed that the optimal NO₃^-/NH₄⁺ ratio was consisted with NO₃^-/NH₄⁺ (nitrate nitrogen/ammonium nitrogen)ratio in B₅ medium. In other words, the suspension culture would be benefitial for the accumulation of chlorogenicacid and growth of cell when NO₃^-/NH₄⁺ (nitrate nitrogen/ ammonium nitrogen)ratio was 13:1. When single N source was added, the biomass of callus reached the maximum(19.26 g·L^-1)in 3.5 g·L^-1KNO₃. However, when KNO₃ concentration was at relatively low levels(0.5 and 1.5 g·L^-1), high chlorogenic acid production was accumulated. The two studies of NO₃^- had some differences from the control concentration(2.5 g·L^-1). What's more, while(NH₄)₂SO₄ concentration(0.268 g·L^-1)exceeded control concentration(0.134 g·L^-1), the highest growth of biomass of the L. macranthoides Hand.-Mazz. “Yulei 1” and the high chlorogenic acid production were observed. In the same way, different species and different concentrations of P, Ca and Mg might have different impacts on the chlorogenic acid content and growth of cell. The result showed that when the highest growth of biomass and the high chlorogenic acid production were accumulated, when NaH₂PO₄·2H₂O concentration was 0.10 g·L^-1. In addition, when the highest growth of biomass and the high chlorogenic acid production were accumulated, when CaCl₂ concentration was 0.20 g·L^-1. So far as Mg²⁺ was concerned, a low MgSO₄·7H₂O concentration could promote the growth of cell, while excessive MgSO₄·7H₂O concentration would promote the accumulation of chlorogenic acid. Taking the biomass growth and chlorogenic acid production into consideration, the optimal concentration of Mg was moderate. These results had some differences from the control. Our study demonstrated that inorganic salt concentration had some differences from inorganic salt concentration of B₅ in suspension cultures of L. macranthoides Hand.-Mazz. “Yulei 1”. This study suggested that the optimized conditions could improve biomass accumulation and chlorogenic acid production through the addition of moderate concentration of inorganic salt in cell suspension cultures of L. macranthoides Hand.-Mazz. “Yulei 1”. These results lays a foundation for large-scale production of chlorogenic acid by using the cell suspension cultures of L. macranthoides Hand.-Mazz. “Yulei 1”. This study also has practical significance to large-scale productions of chlorogenic acid.]]> 2016/5/31 17:34:54 *]]> TANG Ming, WANG Chao, TAN Yun-Ya, LI Qun^* 5true TNFR-Fc in rice seed, we constructed seed-specific expression vector of rice-preferred codon optimized TNFR-Fc (RfTNFR-Fc), by analyzing the whole genomic difference in codon preference and replacing bases with rice-preferred codons. Rice seed specific promoter Glu-4 was amplified via PCR and ligated into pCAMBIA 1381, followed by the ligation with full length synthetic RfTNFR-Fc. The construct was identified by PCR, double enzyme digestion and sequencing. The preferred codons of L, S, P and R differed between rice and human, though most of the amino acids shared the same preferred codons. In the optimized RfTNFR-Fc, 30.8% of the amino acids changed compared with the original version. PCR, double enzyme digestion and sequencing identified that pCAMBIA Glu -RfTNFR-Fc was successfully constructed. This study lays foundation for the expression of TNFR-Fc in rice seed, further facilitating its commercial production.]]> 2016/5/31 17:34:54 *]]> DUAN Yong-Bo, ZHAO Feng-Lan, YAO Lei, TENG Jing-Tong, SHENG Wei, ZHANG Ai-Min, XUE Jian-Ping^* 4true Acacia mangium is a number of Mimosaceae Acacia Mill., and it is planted and promoted in Guangdong, Fujian and Hainan provinces and Guangxi Autonomus Region. A. mangium is used widely, and it is hybrid breeding has received great attention. The research on the A. mangium pollen is key to the hybrid breeding. In this study, we studied the effects of the different sucrose concentrations(50, 100, 150, 200, 250 and 300 g·L^-1), the different boric acid concentrations(100, 200 and 300 mg·L^-1)and different culture temperatures(26, 28 and 30 ℃)on the pollen in vitro germination using the fresh pollen from one plant of A. mangium as experiment materials detected the pollen viability of the other 3 plants of A. mangium using the best A. mangium pollen in vitro germination treatment, whose growths and developments closed to the preliminary A. mangiums and observed and recorded the characteristics of A. mangium pollen in vitro germination. The results showed that the inflorescences of just open of A. mangium were collected and took to the chamber and air dried at room temperature. On the next day after 10:00, a large number of pollen was collected successfully with brush method. The pollen of high purity by microscope detection could ensure the follow-up study in the future. The optimum treatments of pollen germination in vitro of A. mangium were 200 g·L^-1 sucrose, 100 mg·L^-1 boric acid and 28 ℃ culture temperature. After A. mangium pollen cultured for 24 h, the pollen germination percentage was the highest(94.28%)and the pollen tube was the longest(6.0 D). The pollen germination percentage of the other 3 plants of A. mangium had significant difference, 90.11%, 82.31%, 85.67%, respectively. This study provide the basis for further development of artificial control pollination and breeding excellent new hybrid of Acacia.]]> 2016/5/31 17:34:54 *]]> ZHAN Ni, HUANG Lie-Jian^* 3true Brassica pekinensis seeds, Triticum aestivum seeds and Vigna radiate seeds, through the methanol extract the shell and embryo of Euscaphis japonica, with different original extract concentrations of 10%, 20%, 30% and 40% and extracted and separated the endogenous inhibitors in methanol leaching liquid of shell and embryo, the activity and ingredients of endogenous inhibitors in Euscaphis japonicaseeds. The results showed that with the increase of leaching solution concentration, the acid phosphatase activity and germination rate of Brassica pekinensis seeds significantly decreased(P<0.05), the inhibitory effects were showed increasing, and the inhibitory effects of shell were less than that of embryo; the root length and seedling heights promotion in low concentrations and inhibitory in high concentrations, and the maximum at the concentration of 10%, the promotion of shell less than embryo. The amylase activity, germination rate, root length and seedling height of Triticum aestivum seeds decreased gradually(P<0.05), the inhibitory effects were showed increasing gradually, while the germination rate was not significantly different at the concentration less than 20%(P>0.05)and it significantly decreased at 20% concentration(P<0.05), it was 0 at 40% concentration, the inhibitory effects of shell were than that of embryo. The protease activity, germination rate and seedling root length and shoot height of Vigna radiata seeds significantly decreased in the leaching solution concentration higher than 20%(P<0.05), and the effects of shell less than embryo. The results of extraction and separation of endogenous inhibitors showed that there were more phenolic acids and less alkaloids in shell than in embryo. It was concluded that high activating endogenous inhibitors existed in shell and embryo of Euscaphis japonica, which showed differences on properties, composition and content. Endogenous inhibitors in shell were mainly starchy materials, which in embryo mainly affected oil and protein materials.]]> 2016/5/31 17:34:54 *, LI Xi, LIN Rui]]> LIAO Yuan-Lin, CAI Shi-Zhen^*, LI Xi, LIN Rui 2true Panax notoginseng seedlings. The results showed that shoot height, root length, soluble protein content, root activity, CAT and POD activity of P. notoginseng seedlings decreased. Among them, shoot height and POD activity of seedlings were decreased obviously by ferulic acid; and the shoot height of 50, 100 mg·L^-1 P-coumaric acid and 100 mg·L^-1 vanillic acid were 16.19%, 16.67% and 29.29% dramatically lower than that of the control group. The root length of seedlings were also reduced significantly by P-coumaric acid, syringic acid and P-hydroxybenzoic acid, and the root activity of vanillic acid were reduced significantly as well,meanwhile the CAT activity of seedlings were decreased by 10, 50, 100 mg·L^-1 Syringic acid, P-hydroxybenzoic acid and vanillic acid. In addition, the content of chlorophyll of seedlings were also reduced obviously by 1 mg·L^-1 ferulic acid and 100 mg·L^-1 vanillic acid. Furthermore, the content of MDA increased at higher concentrations of ferulic acid, P-coumaric acid, syringic acid and P-hydroxybenzoic acid, while it decreased significantly at 0.1, 1, 10 and 100 mg·L^-1 concentrations of vanillic acid. Besides that, the SOD activity of seedlings increased when treated with syringic acid, vanillic acid, P-hydroxybenzoic acid and higher concentrations of P-coumaric acid, moreover vanillic acid increased them significantly. The results showed that five phenolic acids had some certain allelopathic inhibitory effects on P. notoginseng seedlings, but the effects were not completely consistent. Among them, ferulic acid had greater influence. These provides some theoretical reference for further research of allelopathic autotoxicity of P. notoginseng.]]> 2016/5/31 17:34:54 1, ZHANG Hong-Rui¹, ZHANG Zi-Long^2*, GAO Zhi-Ming¹]]> SHEN Yu-Cong¹, ZHANG Hong-Rui¹, ZHANG Zi-Long^2*, GAO Zhi-Ming¹ 1true