Crops ›› 2018, Vol. 34 ›› Issue (6): 162-167.doi: 10.16035/j.issn.1001-7283.2018.06.026

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The Effects of Different Disinfection Methods on Seed Germination and Study on the Environmental Bacteria in Safflower (Carthamus tinctorius L.)

Ma Mengxue,Zhao Lingling,Tang Si,Chen Xianjun,Qin Rui   

  1. College of Life Sciences, South-Central University for Nationalities/Hubei Provincial Key Laboratory for Protection and Application of Special Plant Germplasm in Wuling Area of China, Wuhan 430074, Hubei, China
  • Received:2018-05-09 Revised:2018-10-11 Online:2018-12-15 Published:2018-12-06

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Abstract:

Contamination was a serious problem in seed germination and tissue culture of safflower (Carthamus tinctorius L.). In our study, seeds were disinfected in order to reduce the contamination rate.Three factors including soaking time, disinfecting agents and disinfecting time were used for L9(3 4) orthogonal experiment. Results were comprehensively evaluated by using the weighted scoring method based on indexes such as germination rate, germination vigor and contamination rate. Orthogonal test showed that the disinfecting agent was the most important factor affecting the seeds germination. Among various treatments, seeds soaked in sterile water for 12h followed by the 2% sodium hypochlorite (NaClO) for 10min was the best disinfecting method in safflower seeds which yield a contamination rate of 8% and a germination rate of 90.67%. In addition, 16S rDNA (GenBank Accession No. MH190221) of a bacterium was isolated from seeds and purified after disinfection which was amplified by PCR. BLAST results showed that it belonged to Bacillus aryabhatti which was most likely to be entophytes in safflower seeds. The results of this study provided an important basis for obtaining well-grown aseptic seedlings, tissue culture as well as genetic transformation.

Key words: Carthamus tinctorius L., Seed disinfection, Seed germination rate, Orthogonal experiment, Bacteria

Table 1

The list of different factors for orthogonal experiment"

水平
Level		 因素Factor
A预浸泡时间(h)
Soaking time		 B消毒剂
Disinfecting agent		 C消毒时间(min)
Disinfecting time		 D对照
Control
1 0 0.1% HgCl2 10 —
2 6 2% NaClO 15 —
3 12 10% H2O2 20 —

Table 2

The effects of orthogonal test of L9(34) to seeds germination rate"

处理
Treatment		 A
预浸泡时间(h)
Soaking time		 B
消毒剂种类
Disinfecting agent		 C
消毒时间(min)
Disinfecting time		 D
对照
Control		 染菌率(%)
Contamination
rate		 发芽势(%)
Germination vigor		 发芽指数(%)
Germination index		 发芽率(%)
Germination
rate		 综合评分
Overall score
T1 0 1（0.1% HgCl2） 10 1 9.33 34.67 33.65 64.67 0.27
T2 0 2（2% NaClO） 15 2 19.33 34.00 38.68 71.33 0.15
T3 0 3（10% H2O2） 20 3 24.00 24.00 28.95 48.67 -0.06
T4 6 1（0.1% HgCl2） 15 3 3.33 50.00 45.37 75.33 0.45
T5 6 2（2% NaClO） 20 1 10.67 55.33 57.22 80.67 0.39
T6 6 3（10% H2O2） 10 2 27.33 36.00 40.02 61.33 -0.01
T7 12 1（0.1% HgCl2） 20 2 2.00 53.33 51.10 85.33 0.53
T8 12 2（2% NaClO） 10 3 8.00 59.33 63.17 90.67 0.48
T9 12 3（10% H2O2） 15 1 18.67 44.67 46.08 67.33 0.17
K1 0.36 1.24 0.74 0.83
K2 0.83 1.02 0.77 0.67
K3 1.18 0.11 0.86 0.88
k1 0.12 0.41 0.25 0.28
k2 0.28 0.34 0.26 0.22
k3 0.39 0.04 0.29 0.29
R 0.28 0.38 0.04 0.07

Fig.1

The effects of different treatments on seed germination rate, germination vigor and germination index"

Table 3

Variance analysis of different factors"

因素Factor 偏差平方和
Square of deviance		 自由度
Degrees of freedom		 F
A预浸泡时间(h)
Soaking time		 0.113 2 37.667*
B消毒剂种类
Disinfecting agent		 0.247 2 82.333*
C消毒时间(min)
Disinfecting time		 0.003 2 1.000
D对照Control 0.000 2

Table 4

Results of verification tests %"

编号
Number		 发芽率
Germination rate		 发芽势
Germination vigor		 染菌率
Contamination rate
1 91.33 60.00 7.33
2 90.67 60.67 7.33
3 90.00 58.00 8.67
[1] 胡尊红, 王沛琦, 杨谨 , 等. 红花分子标记研究进展. 分子植物育种, 2017,15(9):3586-3591.
[2] Ashrafi E, Razmjoo K . Effect of irrigation regimes on oil content and composition of safflower (Carthamus tinctorius L.) cultivars. Journal of the American Oil Chemists’ Society, 2010,87(5):499-506.
doi: 10.1007/s11746-009-1527-8
[3] Lee G A, Sung J S, Lee S Y , et al. Genetic assessment of safflower (Carthamus tinctorius L.) collection with microsatellite markers acquired via pyrosequencing method. Molecular Ecology Resources, 2013,14(1):69-78.
doi: 10.1111/1755-0998.12146 pmid: 23875976
[4] 吴应祥, 黎大爵 . 红花 . 北京: 中国农业出版社, 1982.
[5] 王书云, 李国玉, 黄健 , 等. GC法测定维吾尔药红花籽和红花籽油中亚油酸的含量. 中国药师, 2013,16(1):13-17.
doi: 10.3969/j.issn.1008-049X.2013.01.004
[6] 姜黎 . 红花籽油的功效及应用前景分析. 农产品加工, 2017(12):56-57,60.
doi: 10.16693/j.cnki.1671-9646(X).2017.06.048
[7] 吕培霖, 李成义, 王俊丽 . 红花籽油的研究进展. 中国现代中药, 2016,18(3):387-389.
doi: 10.13313/j.issn.1673-4890.2016.3.030
[8] Velasco L, Perezvich B, Fernandezmartinez J M . Identification and genetic characterization of a safflower mutant with a modified tocopherol profile. Plant Breeding, 2005,124(5):459-463.
doi: 10.1111/j.1439-0523.2005.01150.x
[9] Ilkılıç C, Aydın S, Behcet R , et al. Biodiesel from safflower oil and its application in a diesel engine. Fuel Processing Technology, 2011,92(3):356-362.
doi: 10.1016/j.fuproc.2010.09.028
[10] Mcpherson M A, Yang R C, Good A G , et al. Potential for seed-mediated gene flow in agroecosystems from transgenic safflower (Carthamus tinctorius L.) intended for plant molecular farming. Transgenic Research, 2009,18(2):281-299.
doi: 10.1007/s11248-008-9217-0 pmid: 18941919
[11] Flider F J . Development and commercialization of GLA safflower oil. Lipid Technology, 2013,25(10):227-229.
doi: 10.1002/lite.201300302
[12] Carlsson A S, Zhu L H, Hofvander P , et al. Platform crops amenable to genetic engineering-a requirement for successful production of bio-industrial oils through genetic engineering. Biocatalysis & Agricultural Biotechnology, 2014,3(1):58-64.
doi: 10.1016/j.bcab.2013.12.007
[13] 朱振贤 . 红花的组织培养和品质相关基因的克隆及分析. 南京:南京林业大学, 2011.
[14] 杨捷威, 吴婷婷, 郭美丽 . 红花组织培养的研究进展. 药学服务与研究, 2012,12(1):58-62.
doi: 10.5428/pcar20120117
[15] 丁亦男, 魏健, 张丽娇 . 红花组织培养快繁的研究. 黑龙江畜牧兽医, 2015(2):111-112.
[16] Patial V, Krishna R, Arya G , et al. Development of an efficient,genotype independent plant regeneration and transformation protocol using cotyledonary nodes in safflower (Carthamus tinctorius,L.). Journal of Plant Biochemistry & Biotechnology, 2016,25(4):1-12.
[17] Srinivas B, Luch H, Singh S P , et al. Agrobacterium-mediated transformation of safflower and the efficient recovery of transgenic plants via grafting. Plant Methods, 2011,7(1):12.
doi: 10.1186/1746-4811-7-12 pmid: 21595986
[18] 莫良玉, 吴良欢, 陶勤南 . 植物有机营养研究中供试种子组合灭菌方法比较. 浙江大学学报(农业与生命科学版), 2000,26(6):643-646.
doi: 10.3321/j.issn:1008-9209.2000.06.016
[19] 刘西莉, 牡丽丹, 王红梅 , 等. 红花种子带菌检测及药剂消毒处理. 植物保护, 2003,29(6):49-51.
doi: 10.3969/j.issn.0529-1542.2003.06.017
[20] 李威, 谭勇, 陈文 , 等. 温度对不同品种红花种子萌发的影响. 安徽农业科学, 2013,41(10):4299-4301.
doi: 10.3969/j.issn.0517-6611.2013.10.023
[21] Murashige T, Skoog F . A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 1962,15(3):473-497.
doi: 10.1111/ppl.1962.15.issue-3
[22] 尚丹, 文昭竹, 龚梨霞 , 等. Ca(ClO)2消毒剂对高羊茅种子的消毒效果. 作物研究, 2016,30(5):563-568.
doi: 10.16848/j.cnki.issn.1001-5280.2016.05.19
[23] 江涛, 李晨曦, 陈磊 , 等. 毛竹组织培养中成熟种子消毒方法. 江苏农业科学, 2017,45(4):103-106.
doi: 10.15889/j.issn.1002-1302.2017.04.032
[24] 朱淑颖, 杨帆, 姜叶琴 , 等. 草珊瑚的无菌播种和丛生芽诱导研究. 种子, 2016,35(8):32-36.
doi: 10.16590/j.cnki.1001-4705.2016.08.032
[25] 李威, 张曦, 谭勇 , 等. 干旱胁迫对3个品种红花种子萌发的影响. 中国农学通报, 2013,29(34):207-211.
[26] 朱艳蕾, 安登第, 曾献春 . 银沙槐种子表面消毒技术及无菌萌发条件研究. 新疆师范大学学报(自然科学版), 2014,33(3):17-20.
doi: 10.3969/j.issn.1008-9659.2014.03.004
[27] 李宏, 程平, 郑朝晖 , 等. 盐旱胁迫对3种新疆造林树木种子萌发的影响. 西北植物学报, 2011,31(7):1466-1473.
[28] 谢翔, 顿子云, 刘瑞 , 等. 不同消毒方法及浓度对冰菜种子萌发与幼苗生长的影响. 天津农业科学, 2017,23(6):92-95.
doi: 10.3969/j.issn.1006-6500.2017.06.023
[29] 张晶华, 马亚丽, 兰海燕 . 不同灭菌方法对烟草和拟南芥种子萌发及无菌苗生长的影响. 新疆农业科学, 2016,53(9):1683-1691.
[30] 彭璐, 张志杰, 龚千锋 , 等. 多指标加权法优选醋五倍子炮制工艺. 药物分析杂志, 2017(9):1733-1738.
[31] 胡重怡, 郑少清, 陈兴江 , 等. 烟草无菌苗培养前的种子消毒技术研究. 中国烟草科学, 2007,28(2):45-46.
doi: 10.3969/j.issn.1007-5119.2007.02.012
[32] 张付远, 李丹丹, 廖忠明 , 等. 不同消毒剂及激素对江西省野生白花凤仙花组培诱导的影响. 现代农业科技, 2015(15):164-165.
doi: 10.3969/j.issn.1007-5739.2015.15.098
[33] 陈芳, 党占海, 张建平 , 等. 不同基因型亚麻下胚轴不定芽诱导的研究. 作物杂志, 2014(3):39-43.
[34] 鲁黎明, 安影 . 次氯酸钠对烟草无菌苗生理指标的影响. 作物杂志, 2012(3):141-143.
[35] 倪鑫鑫, 龚海燕, 马顶虹 , 等. 芽孢杆菌生理生化特性分析. 安徽农学通报, 2015(7):47-48.
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