高温和酶解处理水稻秸秆后植物促生物质的鉴定及其效果研究

doi:10.3864/j.issn.0578-1752.2021.15.010

Abstract

Abstract:

【Objective】 To explore the effect of agricultural wastes (rice straw) on plant growth, the identification and analyzation of the small molecules that may exist in the high-temperature extracts and enzymolysis solution of rice straw were performed, and the promoting effect of these substances on cucumber growth was also evaluated.【Method】 The high-temperature extract solution of rice straw was prepared by high temperature water extraction at 115℃ for 30 min. The condensation of the extracellular proteins from Trichoderma guizhouense NJAU 4742 under the induction of rice straw was obtained by ammonium sulfate precipitation method, which was used to hydrolyze the rice straw to prepare rice straw enzymatic hydrolysate. The high-temperature extract and enzymolysis solution of rice straw were diluted into different times, and the growth-promoting effects were verified by cucumber hydroponic experiment. The high-temperature extract and enzymatic hydrolysis of rice straw were identified and compared by UHPLC-QE-MS non-target metabolomics detection technology. Under the control of the acquisition software (Xcalibur 4.0.27, Thermo), the QE mass spectrometer collects primary and secondary mass spectrometry data in the information-related acquisition mode for the high-temperature extract and enzymatic hydrolysate of rice straw. Through the self-written R program package (the kernel was XCMS), the original data was processed for peak identification, peak extraction, peak alignment and integration, and then it was matched with the BiotreeDB (V2.1) self-built MS database for substance annotation. Finally, the growth-promoting effects of some identified substances were verified by the cucumber hydroponic experiment.【Result】 The results showed that both high-temperature extract and enzymatic hydrolysate liquid of rice straw could promote the growth of cucumber seedlings at appropriate concentrations, and the enzymatic hydrolysate of rice straw showed significant growth promoting effect on cucumber when diluted for 100 times. Compared with the CK, the dry weight of overground part, underground part and plant height of cucumber plants treated with 100 times dilution increased by 52.64%, 55.05% and 21.43%, respectively, and the number of plant root tips increased by 31.95%. The high-temperature extract of rice straw owned the best growth promotion effect when it was diluted 50 times, in which the dry weight of overground part, the underground part and the plant height of cucumber plants increased by 44.16%, 63.38% and 55.56%, respectively, and the number of plant root tips increased by 64.44%, compared with CK. The UHPLC-QE-MS non-target detection technology results showed that 714 different substances were identified in the high-temperature extract of rice straw and 638 different substances were identified in the enzymatic hydrolysis solution, among which acetylcholine, L-carnitine and myo-inositol were screened out. Meanwhile, the standard products of these three substances were added to the cucumber root system, and the results showed that they all had considerable promotion effect on cucumber growth. Acetylcholine at the concentration of 1 μmol·L^-1, 10 μmol·L^-1 and 100 μmol·L^-1 could all promote the growth of cucumber. Compared with CK, the exogenous acetylcholine at 1 μmol·L^-1 concentration increases the dry weight of cucumber shoots by 54.69% and the dry weight of roots by 73.67%, the number of root tips increased by 130.5%; Exogenous L-carnitine was beneficial to the growth of cucumber plants at the concentration of 0.1 mmol·L^-1 and 1 mmol·L^-1. Compared with CK, the dry weight of cucumber shoots increased by 33.79% and 30.19%, and the dry weight of roots increased by 44.97% and 48.82%, the number of cucumber root tips increased by 41.8% and 49.9%, respectively; Exogenous myo-inositol at the concentration of 0.05 mmol·L^-1 and 0.1 mmol·L^-1 could promote the growth of cucumber. Compared with CK, the dry weight of cucumber shoots increased by 36.66% and 30.15%, roots dry weight increased by 69.82% and 51.78%, and the number of cucumber root tips increased by 149.0% and 96.7%, respectively.【Conclusion】 In brief, both the high-temperature extract and the enzymatic hydrolysate of rice straw could significantly promote the growth of cucumber, and the acetylcholine, L-carnitine and myo-inositol were detected by LCMS in the pyrolysis and enzymatic hydrolysate of rice straw, which were considered as the functional substances in rice straw.

Key words: rice straw, high-temperature extraction, enzymatic hydrolysis, LCMS, growth promotion

TANG SiYu,LIU QiuMei,MENG XiaoHui,MA Lei,LIU DongYang,HUANG QiWei,SHEN QiRong. Identification of Functional Substances from Rice Straw Obtained by Pyrolysis and Enzymolysis and Its Effect[J].Scientia Agricultura Sinica, 2021, 54(15): 3250-3263.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2021.15.010

https://www.chinaagrisci.com/EN/Y2021/V54/I15/3250

Figures/Tables 10

Fig. 1

Fig. 2

Table 1

Fig. 3

Table 2

Fig. 4

Table 3

Fig. 5

Fig. 6

Table 4

References 33

[1]	WU H S, WANG M Y, ZHOU J, SHI X, HUANG H Y. Potted rice straw increased the antioxidase activity and growth of Chinese green cabbage seedlings and altered the soil microbial community. Archives of Agronomy and Soil Science, 2016, 63(6):784-795. doi: 10.1080/03650340.2016.1235782
[2]	慕兰, 孙笑梅, 王立河, 王喜枝, 姚利娟. 不同秸秆腐熟剂在玉米秸秆粉碎还田中的应用效果研究. 现代农业科技, 2013(16):215-216.
	MU L, SUN X M, WANG L H, WANG X Z, YAO L J. Application effects of different straw decomposing agents in crushing corn stalks and returning to the field. Modern Agricultural Science and Technology, 2013(16):215-216. (in Chinese)
[3]	陈丽芬. 丽水市莲都区秸秆还田技术的探讨. 浙江农业科学, 2018, 59(6):1054-1056.
	CHEN L F. Discussion on straw returning technology in Liandu District, Lishui. Zhejiang Agricultural Science, 2018, 59(6):1054-1056. (in Chinese)
[4]	TIAN Z W, GE Y X, ZHU Q, YU J H, ZHOU Q, CAI J, JIANG D, CAO W X, DAI T B. Soil nitrogen balance and nitrogen utilization of winter wheat affected by straw management and nitrogen application in the Yangtze river basin of China. Archives of Agronomy and Soil Science, 2019, 65(1):1-15. doi: 10.1080/03650340.2018.1479743
[5]	陈捷, 朱洁伟, 张婷, 王秉丽. 木霉菌生物防治作用机理与应用研究进展. 中国生物防治学报, 2011, 27(2):145-151.
	CHEN J, ZHU J W, ZHANG T, WANG B L. Research progress on mechanism and application of biological control of Trichoderma. Chinese Journal of Biological Control, 2011, 27(2):145-151. (in Chinese)
[6]	尹婷, 徐秉良, 梁巧兰, 古丽君, 李荣峰. 耐药性木霉T2菌株的筛选、紫外诱变与药剂驯化. 草业学报, 2013, 22(2):117-122.
	YIN T, XU B L, LIANG Q L, GU L J, LI R F. Screening, UV mutagenesis and drug domestication of drug-resistant Trichoderma T2 strain. Journal of Prataculture, 2013, 22(2):117-122. (in Chinese)
[7]	杨春林, 席亚东, 刘波微, 张敏, 彭化贤. 哈茨木霉T-h-30对几种蔬菜的促生作用及病害防治初探. 西南农业学报, 2008, 21(6):1603-1607.
	YANG C L, XI Y D, LIU B W, ZHANG M, PENG H X. Growth promoting effect of Trichoderma harzianum T-h-30 on several vegetables and disease control. Southwest China Journal of Agricultural Sciences, 2008, 21(6):1603-1607. (in Chinese)
[8]	冯程龙, 王晓婷, 康文晶, 孟晓慧, 张风革, 冉炜, 沈其荣. 利用小麦秸秆生产木霉分生孢子及其生物有机肥对黄瓜的促生效果. 植物营养与肥料学报, 2017, 3(5):1286-1295.
	FENG C L, WANG X T, KANG W J, MENG X H, ZHANG F G, RAN W, SHEN Q R. Growth promoting effect of Trichoderma conidia and its bioorganic fertilizer on cucumber using wheat straw. Journal of Plant Nutrition and Fertilizers, 2017, 3(5):1286-1295. (in Chinese )
[9]	郎剑锋, 刘起丽, 杨蕊, 石明旺, 陈锡岭. 利用秸秆和肥料增殖木霉及对玉米茎基腐病的生物防治. 玉米科学, 2019, 27(2):161-169.
	LANG J F, LIU Q L, YANG R, SHI M W, CHEN X L. Biocontrol of Trichoderma with straw and fertilizer on maize stem basal rot disease. Journal of Maize Sciences, 2019, 27(2):161-169. (in Chinese)
[10]	STOTTMEISTER U. Altlastsanierung mit Huminstoffsystemen. Prinzipien der natur in der Umwelttechnologie. Chemie in Unserer Zeit, 2008, 42(1):24-41. doi: 10.1002/(ISSN)1521-3781
[11]	GAGNON R, LEVASSEUR M, WEISE A M, FAUCHOT J, CAMPBELL P G. C, WEISSENBOECK B J, MERZOUK A, GOSSELIN M. Growth stimulation of Alexandrium tamarense (dinophyceae) by humic substances from the Manicouagan River (eastern Canada). Journal of Phycology, 2010, 41(3):489-497. doi: 10.1111/(ISSN)1529-8817
[12]	JINDO K, MARTIM S A, NAVARRO E C, PEREZ-ALFOCEA F, HERNANDEZ T, GARCIA C, AGUIAR N O, CANELLAS L P. Root growth promotion by humic acids from composted and non- composted urban organic wastes. Plant and Soil, 2012, 353(1/2):209-220. doi: 10.1007/s11104-011-1024-3
[13]	GAO J K, LIANG C L, SHEN G Z, LV J L, WU H M. Spectral characteristics of dissolved organic matter in various agricultural soils throughout China. Chemosphere, 2017, 176(6):108-116. doi: 10.1016/j.chemosphere.2017.02.104
[14]	TREVISAN S, FRANCIOSO O, QUAGGIOTTI S, NARDI S. Humic substances biological activity at the plant-soil interface. Plant Signaling & Behavior, 2010, 5(6):635-643.
[15]	MA X, HU J, WANG X, CHO S, GAO M. T. An integrated strategy for the utilization of rice straw: Production of plant growth promoter followed by ethanol fermentation. Process Safety and Environmental Protection, 2019, 129(9):1-7. doi: 10.1016/j.psep.2019.06.004
[16]	杨玉春. 介绍一种简易测定黄瓜叶面积的方法. 辽宁农业科学, 1981(1):42-43.
	YANG Y C. Introduction of a simple method to determine the leaf area of cucumber. Liaoning Agricultural Sciences, 1981(1):42-43. (in Chinese)
[17]	ISIK N, ALTEHELD B, KUHN S, SCHULZE-KAYSERS N, KUNZ B, WOLLSEIFEN H R, STEHLE P, LESSER S. Polyphenol release from protein and polysaccharide embedded plant extracts during in vitro digestion. Food Research International, 2014, 65:109-114. doi: 10.1016/j.foodres.2014.02.012
[18]	冯程龙. 哈茨木霉T-E5分生孢子固体发酵工艺及其生物有机肥对黄瓜的促生效果研究[D]. 南京: 南京农业大学, 2017.
	FENG C L. Study on solid-state fermentation of Trichoderma harzianum T-E5 conidia and growth promoting effect of its bio-organic fertilizer on cucumber[D]. Nanjing: Nanjing Agricultural University, 2017. (in Chinese)
[19]	JIA W, COLLINS S R A, ADAM E, NIKOLAUS W, JO D, ROBERTS I N, WALDRON K W. Release of cell wall phenolic esters during hydrothermal pretreatment of rice husk and rice straw. Biotechnology for Biofuels, 2018, 11(1):162. doi: 10.1186/s13068-018-1157-1
[20]	CHEN Y, HUANG J, LI Y, ZENG G, ZHOU W. Study of the rice straw biodegradation in mixed culture of Trichoderma viride and Aspergillus niger by gc-ms and ftir. Environmental Science and Pollution Research, 2015, 22(13):9807-9815. doi: 10.1007/s11356-015-4149-8
[21]	JUNG D U, YOO H Y, KIM S B, LEE J H, PARK C, KIM S W. Optimization of medium composition for enhanced cellulase production by mutant Penicillium brasilianum KUEB15 using statistical method. Journal of Industrial & Engineering Chemistry, 2015, 25:145-150.
[22]	LIAO H P, FAN X T, MEI X L, WEI Z, RAZA W, SHEN Q R, XU Y C. Production and characterization of cellulolytic enzyme from Penicillium oxalicum GZ-2 and its application in lignocellulose saccharification. Biomass & Bioenergy, 2015, 74(3):122-134.
[23]	ELLILA S, FONSECA L, UCHIMA C, COTA J, GOLDMAN G H, SALOHEIMO M, SACON V, SIIKA-AHO M. Development of a low-cost cellulase production process using Trichoderma reesei for Brazilian biorefineries. Biotechnology for Biofuels, 2017, 10(1):30. doi: 10.1186/s13068-017-0717-0
[24]	BAMEL K, GUPTA S C, GIPTA R. Acetylcholine causes rooting in leaf explants of in vitro raised tomato (Lycopersicon esculentum Miller) seedlings. Life Sciences, 2007, 80(24/25):2393-2396. doi: 10.1016/j.lfs.2007.01.039
[25]	AURELIE C, RIPPA S, AGNES Y, NGUYEN P J, RENOU J P, PERRIN Y. The effect of carnitine on Arabidopsis development and recovery in salt stress conditions. Planta, 2012, 235(1):123-135. doi: 10.1007/s00425-011-1499-4
[26]	AYTUN Y, CEVIK S, UNYAYAR S. Effects of exogenous myo-inositol on leaf water status and oxidative stress of Capsicum annuum under drought stress. Acta Physiologiae Plantarum, 2018, 40(6):122. doi: 10.1007/s11738-018-2690-z
[27]	ROSHCHINA V V. Neurotransmitters in plant life. Phytochemistry, 2001, 63(3):373-373. doi: 10.1016/S0031-9422(03)00144-4
[28]	BAMEL K, GUPTA R, GUPTA S C. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitroraised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby. Plant Signaling & Behavior, 2016, 11(6):e1187355.
[29]	PEKALA J, PATKOWSKA-SOKOLA B, BODKOWSKI R, JAMROZ D, NOWAKOWSKI P, LOCHYNSKI S, LIBROWSK T. L-carnitine metabolic functions and meaning in humans life. Current Drug Metabolism, 2011, 12(7):667-678. doi: 10.2174/138920011796504536
[30]	RIPPA S, ZHAO Y, MERLIER F, AURELIEC, PERRIN Y. The carnitine biosynthetic pathway in Arabidopsis thaliana shares similar features with the pathway of mammals and fungi. Plant Physiology & Biochemistry, 2012, 60(3):109-114.
[31]	ONEY-BIROL S. Exogenous L-carnitine promotes plant growth and cell division by mitigating genotoxic damage of salt stress. Scientific Reports, 2019, 9(2):708-716. doi: 10.1038/s41598-018-37516-4
[32]	ENGLE D B, BELISLE J A, GUBBELS J A A, PETRIE S E, HUTSON P R, KUSHNER D M, PATANKAR M S. Effect of acetyl-l-carnitine on ovarian cancer cells' proliferation, nerve growth factor receptor (Trk-A and p75) expression, and the cytotoxic potential of paclitaxel and carboplatin. Gynecologic Oncology, 2009, 112(3):631-636. doi: 10.1016/j.ygyno.2008.11.020
[33]	HU L Y, ZHOU K, LI Y T S, CHEN X F, LIU B B, LI C Y, GONG X Q, MA F W. Exogenous myo-inositol alleviates salinity-induced stress in Malus hupehensis Rehd. Plant Physiology and Biochemistry, 2018, 133:116-126. doi: 10.1016/j.plaphy.2018.10.037

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

处理 Treatment	茎粗 Stem diameter (mm)	株高 Plant height (cm)	根系指标Root index
处理 Treatment	茎粗 Stem diameter (mm)	株高 Plant height (cm)	总根长 Total roots length (cm)	根表面积 Root surface area (cm²)	根尖数 Root tips
CK	4.20±0.20d	11.2±0.5c	1046.8±69.4b	162.71±5.34c	1111±114b
25	4.47±0.17c	11.6±0.4c	774.1±117.9c	128.97±8.78a	1124±239b
50	4.88±0.13b	13.0±0.6ab	1153.1±87.6b	194.44±11.99ab	1483±160a
100	5.14±0.06a	13.6±0.8a	1391.9±121.7a	204.39±15.17a	1466±82a
200	4.29±0.06cd	12.6±0.3b	1060.4±60.0b	177.11±10.74bc	1115±76b

处理 Treatment	茎粗 Stem diameter (mm)	株高 Plant height (cm)	根系指标Root index
处理 Treatment	茎粗 Stem diameter (mm)	株高 Plant height (cm)	总根长 Total roots length (cm)	根表面积 Root surface area (cm²)	根尖数 Root tips
CK	3.88±0.05b	10.8±0.3c	909.4±20.8bc	132.07±5.25c	1094±14e
200	3.91±0.10b	13.2±0.6b	869.9±18.4c	135.40±10.94c	1207±53d
100	3.94±0.10b	13.5±0.4b	938.1±30.1b	154.23±6.95b	1463±29c
50	4.41±0.09a	16.8±0.4a	1225.4±48.0a	190.00±9.39a	1799±51a
25	0.11±0.11a	16.8±0.3a	1172.3±43.0a	187.81±3.26a	1670±56b

物质名称 Substance name	二级质谱打分值 MS2 score	保留时间 Retention time (s)	质荷比 Charge-to- mass ratio	绝对峰面积平均值 Average absolute peak area
物质名称 Substance name	二级质谱打分值 MS2 score	保留时间 Retention time (s)	质荷比 Charge-to- mass ratio	灭活对照 CK	酶解液 Enzymatic hydrolysate	高温浸提液 High temperature extract
乙酰胆碱 Acetylcholine	0.82	351.71	146.12	0.00	1232294.50	1304286316.78
左旋肉碱 L-Carnitine	0.99	348.51	162.11	0.00	1057738.90	974337335.47
肌醇 myo-Inositol	0.92	387.48	203.05	2533514.18	438705294.96	35929119.59

物质名称 Substance name	浓度 Concentration	新叶面积 Leaf area (cm²)	新叶叶绿素 SPAD value	茎粗 Stem diameter (mm)	根系指标Root index
物质名称 Substance name	浓度 Concentration	新叶面积 Leaf area (cm²)	新叶叶绿素 SPAD value	茎粗 Stem diameter (mm)	总根长 Total roots length (cm)	根表面积 Root surface area (cm²)	根尖数 Root tips
氯化乙酰胆碱 Acetylcholine chloride	CK	17.90±1.20b	35.9±1.1c	3.28±0.04c	584.3±24.8b	98.65±4.91b	727±70c
	1 μmol·L^-1	36.5±0.60a	46.5±0.8a	3.82±0.02ab	901.7±25.6a	133.54±3.32a	1676±29a
	10 μmol·L^-1	36.41±2.26a	44.9±0.3b	3.89±0.05a	869.9±90.5a	125.78±10.33a	1643±107a
	100 μmol·L^-1	34.98±3.30a	44.1±0.3b	3.74±0.06b	882.4±81.6a	127.03±7.55a	1313±126b
左旋肉碱 L-Carnitine	CK	17.90±1.20b	35.9±1.1c	3.28±0.04c	584.3±24.8b	98.65±4.91b	727±70c
	0.1 mmol·L^-1	25.10±1.47a	39.2±1.0b	3.70±0.06a	686.5±63.5a	118.38±3.93a	1031±45a
	1 mmol·L^-1	22.17±3.34a	41.2±0.7a	3.68±0.01a	744.±45.8a	109.87±3.78a	1090±21a
	5 mmol·L^-1	22.11±1.95a	42.6±0.5a	3.53±0.03b	520.6±41.5b	86.77±7.34c	896±67b
肌醇 myo-Inositol	CK	17.90±1.20b	35.9±1.1c	3.28±0.04b	584.3±24.8c	98.65±4.91c	727±70c
	0.05 mmol·L^-1	29.79±3.09a	44.0±0.4a	3.95±0.01a	1026.8±74.1a	138.27±5.83a	1810±186a
	0.1 mmol·L^-1	27.13±3.10a	44.5±0.2a	3.82±0.11a	808.4±18.3b	112.23±9.55b	1430±125b
	1 mmol·L^-1	-	-	2.86±0.15c	102.8±18.3d	18.96±6.23d	477±93d

Identification of Functional Substances from Rice Straw Obtained by Pyrolysis and Enzymolysis and Its Effect

RichHTML

PDF

Abstract

Cite this article

share this article

Figures/Tables 10

References 33

Related Articles 15

Metrics

Comments

Recommended 0

[1]	LIU ShuJun,LI DongChu,HUANG Jing,LIU LiSheng,WU Ding,LI ZhaoQuan,WU YuanFan,ZHANG HuiMin. Effects of Straw Returning and Potassium Fertilizer on Soil Aggregate and Potassium Distribution Under Rapeseed-Rice Rotation [J]. Scientia Agricultura Sinica, 2022, 55(23): 4651-4663.
[2]	LI Yu,WANG Fang,WENG ZeBin,SONG HaiZhao,SHEN XinChun. Preparation of Soybean Protein-Derived Pro-osteogenic Peptides via Enzymatic Hydrolysis [J]. Scientia Agricultura Sinica, 2021, 54(13): 2885-2894.
[3]	Ke YU,Lei LIU,RuiFen ZHANG,JianWei CHI,XuChao JIA,MingWei ZHANG. Effect of Pre-Enzymatic-Drum Drying Process on the Quality of Whole Wheat Flakes [J]. Scientia Agricultura Sinica, 2020, 53(6): 1256-1268.
[4]	Lu YANG,NaoHua ZENG,JinShun BAI,Xing ZHOU,GuoPeng ZHOU,SongJuan GAO,Jun NIE,WeiDong CAO. Responses of Soil Diazotroph Community to Rice Straw, Glucose and Nitrogen Addition During Chinese Milk Vetch Growth [J]. Scientia Agricultura Sinica, 2020, 53(1): 105-116.
[5]	BAI RuXia,ZENG HuiWen,FAN Qian,YIN Jie,SUI ZongMing,YUAN Ling. Effects of Ceriporia lacerata on Gummy Stem Blight Control, Growth Promotion and Yield Increase of Cucumbers [J]. Scientia Agricultura Sinica, 2019, 52(15): 2604-2615.
[6]	QIU CunPu, CHEN XiaoFen, LIU Ming, LI WeiTao, WU Meng, JIANG ChunYu, FENG YouZhi, LI ZhongPei. Microbial Transformation Process of Straw-Derived C in Two Typical Paddy Soils [J]. Scientia Agricultura Sinica, 2019, 52(13): 2268-2279.
[7]	CHEN Si-yuan, LIU Yong-xiang, CAO Xiao-zhou, ZHANG Yi-jing, CHEN Hai-juan, SHEN Xin-chun. The Preparation Process for Isolation of a Highly Active Antioxidant Peptide Derived from Wheat Germ Albumin [J]. Scientia Agricultura Sinica, 2016, 49(12): 2379-2388.
[8]	WEN Wei, LIU Lei, ZHANG Ming-wei, ZHANG Rui-fen, WEI Zhen-cheng, TANG Xiao-jun, MA Yong-xuan. The Optimal Composite Enzymatic Hydrolysis Process on Defatted Rice Bran and Its Nutritional Evaluation [J]. Scientia Agricultura Sinica, 2015, 48(8): 1597-1608.
[9]	HUANG Jing, GU Ming-Hua, XU Shi-Hong, YANG Wei-Fang, JIANG Li-Geng. Effects of No-Tillage and Rice-Seedling Casting with Rice Straw Returning on Content of Nitrogen, Phosphorus and Potassium of Soil Profiles [J]. Scientia Agricultura Sinica, 2012, 45(13): 2648-2657.
[10]	DI Xiu-Cai, LIU Ming, LI Zhong-Pei, XU Yang-Chun. Effect of Different Additives on Decomposition of Rice Straw [J]. Scientia Agricultura Sinica, 2012, 45(12): 2412-2419.
[11]	LIU Qing-li ,SHI Jun-xiong,ZHANG Yun-gui ,WANG Jing-jun,LI Zhi-hong . The Effects of Various Organic Matters on the Nitrogen Nutrition of Flue-Cured Tobacco and Its Quality by 15N [J]. Scientia Agricultura Sinica, 2010, 43(22): 4642-4651 .
[12]	. Study on Optimal Hydrolysis Process for Preparing Rice Bran Short Peptides with Two Enzymes [J]. Scientia Agricultura Sinica, 2009, 42(5): 1744-1750 .
[13]	,,,,. Effects of Soil Moisture Content on the Mineralization of Added 14C-Labbelled Straw and Native Soil Organic Carbon in Upland Soil [J]. Scientia Agricultura Sinica, 2006, 39(03): 538-543 .
[14]	,,. Enzymatic Preparation and Functional Properties of Wheat Gluten Hydrolysates [J]. Scientia Agricultura Sinica, 2006, 39(03): 593-598 .
[15]	,,,,,,. Effects of Bio-Treatments on Desilicification for Improving Rice Straw Degradability [J]. Scientia Agricultura Sinica, 2006, 39(02): 406-411 .