新疆农业科学 ›› 2023, Vol. 60 ›› Issue (6): 1499-1505.DOI: 10.6048/j.issn.1001-4330.2023.06.024
付强1(), 王琴2, 徐彩芹3, 牛俊丽3, 牛建新1, 王建友3()
收稿日期:
2022-10-19
出版日期:
2023-06-20
发布日期:
2023-06-20
通信作者:
王建友(1964-),男,甘肃兰州人,研究员,硕士生导师,研究方向为经济林,(E-mail) almonds@126.com
作者简介:
付强(1997-),男,内蒙古人,硕士研究生,研究方向为果树栽培生理,(E-mail)fuqiang9777@126.com
基金资助:
FU Qiang1(), WANG Qin2, XU Caiqin3, NIU Junli3, NIU Jianxin1, WANG Jianyou3()
Received:
2022-10-19
Online:
2023-06-20
Published:
2023-06-20
Correspondence author:
WANG Jianyou(1964-), male,researcher,Master's Supervisor,research direction:economic forest, (E-mail) almonds@126.com
Supported by:
摘要:
【目的】筛选出消毒的最佳时间和抗褐化剂的最适浓度,研究外植体细胞结构在褐化过程中的变化,为黑核桃的组培研究和工厂化育苗提供理论基础。【方法】采用不同时长对黑核桃叶片和茎段进行消毒处理,观察其对污染率、褐化率的影响;以AC、PVP、VC 3种抗褐化剂进行正交试验,研究其对黑核桃茎段褐化率和萌发率的影响;观察褐化外植体徒手切片。【结果】 (1)叶片以0.1%HgCl2消毒5 min,污染率和褐化率最低;(2)茎段以二次消毒法,0.1%HgCl2加吐温80消毒5 min,污染率和褐化率最低;(3)以AC 2 g/L、PVP 110 mg/L、VC 100 mg/L处理的褐化率最低;以AC 1 g/L、PVP 70 mg/L、VC 50 mg/L处理的萌发率最高。【结论】叶片以0.1%HgCl2消毒5min为最佳消毒时间;茎段以二次消毒法,0.1%HgCl2加吐温80消毒5 min为最佳消毒时间;最适的抗褐化剂浓度配比为AC1g/L、PVP 110 mg/L、VC100 mg/L。
中图分类号:
付强, 王琴, 徐彩芹, 牛俊丽, 牛建新, 王建友. 黑核桃组培中消毒及防止褐化最适浓度分析[J]. 新疆农业科学, 2023, 60(6): 1499-1505.
FU Qiang, WANG Qin, XU Caiqin, NIU Junli, NIU Jianxin, WANG Jianyou. Study on disinfection and prevention of browning in tissue culture of black walnut[J]. Xinjiang Agricultural Sciences, 2023, 60(6): 1499-1505.
水平 Levels | 抗褐化剂 Anti browning agent | ||
---|---|---|---|
AC (g/L) | PVP (mg/L) | VC (mg/L) | |
1 | 1 | 70 | 50 |
2 | 1.5 | 90 | 100 |
3 | 2 | 110 | 150 |
表1 抗褐化剂L9(34)正交试验因素水平
Tab.1 Factor level of orthogonal experiment of anti browning agent L9 (34)
水平 Levels | 抗褐化剂 Anti browning agent | ||
---|---|---|---|
AC (g/L) | PVP (mg/L) | VC (mg/L) | |
1 | 1 | 70 | 50 |
2 | 1.5 | 90 | 100 |
3 | 2 | 110 | 150 |
处理 Treatment | 消毒试剂 Disinfection reagent | 消毒时长 Disinfection time (min) | 污染率 Pollution rate (%) | 褐化率 Browning rate (%) |
---|---|---|---|---|
A1 | 0.1%HgCl2 | 4 | 22.22a | 13.89b |
A2 | 0.1%HgCl2 | 5 | 11.76ab | 14.71b |
A3 | 0.1%HgCl2 | 6 | 11.43ab | 25.71ab |
A4 | 0.1%HgCl2 | 7 | 8.11b | 37.84a |
表2 不同消毒时间下叶片污染率和褐化率变化
Tab.2 Effects of disinfection time on leaf contamination rate and browning rate
处理 Treatment | 消毒试剂 Disinfection reagent | 消毒时长 Disinfection time (min) | 污染率 Pollution rate (%) | 褐化率 Browning rate (%) |
---|---|---|---|---|
A1 | 0.1%HgCl2 | 4 | 22.22a | 13.89b |
A2 | 0.1%HgCl2 | 5 | 11.76ab | 14.71b |
A3 | 0.1%HgCl2 | 6 | 11.43ab | 25.71ab |
A4 | 0.1%HgCl2 | 7 | 8.11b | 37.84a |
图1 健康外植体与褐化外植体对比 注:A.接种5 d后的外植体;B.腋芽萌动的外植体;C.腋芽萌发的外植体;D.褐化死亡的外植体
Fig.1 Comparison between healthy explants and browning explants Note: A. explants 5 days after inoculation.B. Explants of axillary bud sprouting. C. Explants for axillary bud germination. D. Browning dead explants
处理 Treatment | 消毒时间 Disinfection time (min) | 消毒次数 Disinfection times | 污染率 Pollution rate (%) | 褐化率 Browning rate (%) |
---|---|---|---|---|
B1 | 8 | 单次 | 48.48a | 24.24c |
B2 | 10 | 单次 | 33.33b | 45.45b |
B3 | 12 | 单次 | 14.29c | 57.14a |
B4 | 4 | 二次 | 47.22a | 11.11d |
B5 | 5 | 二次 | 29.41b | 17.65cd |
B6 | 6 | 二次 | 20.59c | 26.67c |
表3 不同消毒时间下茎段污染率和褐化率变化
Tab.3 Effects of different disinfection time on pollution rate and browning rate of stem segments
处理 Treatment | 消毒时间 Disinfection time (min) | 消毒次数 Disinfection times | 污染率 Pollution rate (%) | 褐化率 Browning rate (%) |
---|---|---|---|---|
B1 | 8 | 单次 | 48.48a | 24.24c |
B2 | 10 | 单次 | 33.33b | 45.45b |
B3 | 12 | 单次 | 14.29c | 57.14a |
B4 | 4 | 二次 | 47.22a | 11.11d |
B5 | 5 | 二次 | 29.41b | 17.65cd |
B6 | 6 | 二次 | 20.59c | 26.67c |
处理 Treatment | 抗褐化剂 Anti browning agent | 褐化率 Browning rate (%) | 萌发率 Germination rate (%) | ||
---|---|---|---|---|---|
AC (g/L) | PVP (mg/L) | VC (mg/L) | |||
C1 | 1 | 70 | 50 | 35.00d | 45.00a |
C2 | 1 | 90 | 100 | 23.33e | 41.607ab |
C3 | 1 | 110 | 150 | 38.33cd | 38.33bc |
C4 | 1.5 | 70 | 100 | 56.67a | 33.33cd |
C5 | 1.5 | 90 | 150 | 53.33a | 36.67bcd |
C6 | 1.5 | 110 | 50 | 46.67b | 31.67de |
C7 | 2 | 70 | 150 | 38.33cd | 36.67bcd |
C8 | 2 | 90 | 50 | 40.00cd | 38.33bc |
C9 | 2 | 110 | 100 | 21.67e | 26.67e |
表4 抗褐化剂不同浓度配比下褐化率和萌发率变化
Tab.4 Effects of different concentrations of anti browning agent on browning rate and germination rate
处理 Treatment | 抗褐化剂 Anti browning agent | 褐化率 Browning rate (%) | 萌发率 Germination rate (%) | ||
---|---|---|---|---|---|
AC (g/L) | PVP (mg/L) | VC (mg/L) | |||
C1 | 1 | 70 | 50 | 35.00d | 45.00a |
C2 | 1 | 90 | 100 | 23.33e | 41.607ab |
C3 | 1 | 110 | 150 | 38.33cd | 38.33bc |
C4 | 1.5 | 70 | 100 | 56.67a | 33.33cd |
C5 | 1.5 | 90 | 150 | 53.33a | 36.67bcd |
C6 | 1.5 | 110 | 50 | 46.67b | 31.67de |
C7 | 2 | 70 | 150 | 38.33cd | 36.67bcd |
C8 | 2 | 90 | 50 | 40.00cd | 38.33bc |
C9 | 2 | 110 | 100 | 21.67e | 26.67e |
项目 Items | 抗褐化剂Anti browning agent | 影响主次 Influence order | 最优方案 Optimal scheme | |||
---|---|---|---|---|---|---|
AC(g/L) | PVP(mg/L) | VC(mg/L) | ||||
褐化率 Browning rate | K1 | 96.7 | 130 | 121.7 | AC>VC >PVP | AC 1 g/L、 PVP 110 mg/L、 VC 100 mg/L |
K2 | 156.7 | 116..7 | 101.7 | |||
K3 | 100 | 106.7 | 129.9 | |||
k1 | 32.2 | 43.3 | 40.6 | |||
k2 | 52.2 | 38.9 | 33.9 | |||
k3 | 33.3 | 35.6 | 43.3 | |||
R | 20 | 7.8 | 9.4 | |||
萌发率 Germination rate | K1 | 125.0 | 115.0 | 115.0 | AC>PVP >VC | AC 1 g/L、 PVP 190 mg/L、 VC 50 mg/L |
K2 | 101.7 | 116.7 | 101.7 | |||
K3 | 101.7 | 96.7 | 111.7 | |||
k1 | 41.7 | 38.3 | 38.3 | |||
k2 | 33.9 | 38.9 | 33.9 | |||
k3 | 33.9 | 32.2 | 37.2 | |||
R | 7.8 | 6.7 | 4.4 |
表5 极差分析
Tab.5 Extreme difference analysis
项目 Items | 抗褐化剂Anti browning agent | 影响主次 Influence order | 最优方案 Optimal scheme | |||
---|---|---|---|---|---|---|
AC(g/L) | PVP(mg/L) | VC(mg/L) | ||||
褐化率 Browning rate | K1 | 96.7 | 130 | 121.7 | AC>VC >PVP | AC 1 g/L、 PVP 110 mg/L、 VC 100 mg/L |
K2 | 156.7 | 116..7 | 101.7 | |||
K3 | 100 | 106.7 | 129.9 | |||
k1 | 32.2 | 43.3 | 40.6 | |||
k2 | 52.2 | 38.9 | 33.9 | |||
k3 | 33.3 | 35.6 | 43.3 | |||
R | 20 | 7.8 | 9.4 | |||
萌发率 Germination rate | K1 | 125.0 | 115.0 | 115.0 | AC>PVP >VC | AC 1 g/L、 PVP 190 mg/L、 VC 50 mg/L |
K2 | 101.7 | 116.7 | 101.7 | |||
K3 | 101.7 | 96.7 | 111.7 | |||
k1 | 41.7 | 38.3 | 38.3 | |||
k2 | 33.9 | 38.9 | 33.9 | |||
k3 | 33.9 | 32.2 | 37.2 | |||
R | 7.8 | 6.7 | 4.4 |
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