Study on Callus Induction and Anti-browning of Rosa roxburghii Tratt Leaves

doi:10.11924/j.issn.1000-6850.casb2022-0016

Abstract

Abstract:

In order to further optimize the callus induction conditions of Rosa roxburghii Tratt leaves, the leaves of four genotypes of Rosa roxburghii Tratt, ‘Guinong 1’, ‘Guinong 2’, ‘Guinong 5’ and ‘Guinong 7’, were used as test materials. The effects of pretreatment, sampling time, plant growth regulator and anti-browning agent on leaf callus induction and browning rate reducing were studied by tissue culture method. The results showed that being soaked in 100 mg/LVc for 12 h could significantly increase the induction rate of the four genotypes’ leaves and reduce the browning rate, among which, ‘Guinong 1’ had the highest induction rate of 97.44 %. Spring was the best time for selecting leaves of different genotypes, and the callus induction rate was the highest and the browning rate was the lowest in March. The suitable combination of plant growth regulators for callus induction of ‘Guinong 1’ and ‘Guinong 2’leaves was 1.0 mg/L6-BA+0.5 mg/LNAA, while the optimal combination of plant growth regulators for callus induction from leaves of ‘Guinong 5’ and ‘Guinong 7’ was 1.0 mg/L6-BA+1.0 mg/LNAA+1.0 mg/LTDZ. The effect of three anti-browning agents on callus proliferation and culture of Rosa roxburghii Tratt leaves of the four genotypes was AgNO₃ > citric acid > PVP. Citric acid or AgNO₃ supplementation could significantly reduce the browning rate of Rosa roxburghii Tratt leaves, and 2.0 g/L citric acid or 0.4 g/LAgNO₃was the most suitable, under which, the browning rate of leaves of the four genotypes reached the lowest value. In this experiment, we obtained the most suitable combination of growth regulators for callus induction of the four genotypes of Rosa roxburghii Tratt, which could reduce the browning rate. The study lays a foundation for the establishment of subsequent in vitro regeneration system.

Key words: Rosa roxburghii Tratt, leaves, callus, induction, browning

CLC Number:

S661.9

LI Tingting, CHEN Hong. Study on Callus Induction and Anti-browning of Rosa roxburghii Tratt Leaves[J]. Chinese Agricultural Science Bulletin, 2022, 38(32): 12-21.

Figures/Tables 12

References 26

[1]	刘金萍, 车代弟, 龚束芳, 等. 丰花月季体细胞无性系变异的研究[J]. 东北农业大学学报, 2008, 52(7):17-20.
[2]	王茜龄. 桑杂种胚器官化学诱变与离体再生四倍体育种新材料研究[D]. 重庆: 西南大学, 2006.
[3]	王平, 唐小浪, 马培恰, 等. 辐射诱变和芽变柑橘品种(系)的AFLP分析[J]. 果树学报, 2012, 29(1):130-134.
[4]	邓仁菊, 范建新, 王永清, 等. 火龙果幼苗离体辐射诱变及其对低温胁迫的生理响应[J]. 热带作物学报, 2018, 39(2):260-266.
[5]	王爱连, 王平平, 梁东, 等. 阔叶猕猴桃GalDH基因植物表达载体的构建及转化番茄的研究[J]. 西北农业学报, 2012, 21(1):130-135.
[6]	杜小丽, 陈秀孔, 张计育, 等. 苹果属植物再生体系优化及长富6号遗传转化体系的建立[J]. 江苏农业学报, 2012, 28(3):643-647.
[7]	张绿萍, 陈红. 影响刺梨花药愈伤组织诱导因素的研究[J]. 山地农业生物学报, 2009, 28(1):18-23.
[8]	陈红, 张绿萍. 不同处理对刺梨未受精胚珠愈伤组织诱导的影响[J]. 江西农业学报, 2013, 25(6):49-50,54.
[9]	卢薇, 费建军, 沈晓梅, 等. 五种果汁的酚类组成及其抗氧化、抗肿瘤细胞增殖活性研究[J]. 食品工业科技, 2021, 43(8):1-11.
[10]	连紫凌, 王永, 刘阳, 等. 刺梨资源主要有效成分含量测定及综合质量评价[J]. 中药材, 2021, 44(8):1915-1922.
[11]	房洪舟, 鲁敏, 安华明. 刺梨叶片愈伤组织培养体系建立及其主要活性物质分析[J]. 植物生理学报, 2019, 55(8):1147-1155.
[12]	刘彦珍, 郜新强, 张元臣, 等. 植物激素和光暗培养条件对卷丹百合再生体系的影响[J]. 南方农业学报, 2021, 52(5):1300-1309.
[13]	鲁娇娇, 张梦迪, 孙红梅. 朱顶红花孔雀和黑天鹅离体快繁技术体系的建立[J]. 沈阳农业大学学报, 2020, 51(4):488-493.
[14]	邓仁菊, 范建新, 王永清, 等. 成年火龙果植株茎段愈伤组织诱导、增殖及分化研究[J]. 西南农业学报, 2019, 32(3):595-603.
[15]	张旭红, 王頔, 梁振旭, 等. 欧洲百合愈伤组织诱导及植株再生体系的建立[J]. 植物学报, 2018, 53(6):840-847. doi: 10.11983/CBB17208
[16]	GUO B, ABBASI B H, ZEB A, et al. Thidiazuron:a multi-dimensional plant growth regulator[J]. African journal of biotechnology, 2011, 10(45):8984-9000. doi: 10.5897/AJB11.636 URL
[17]	MURTHY B N S, MURCH S J, SAXENA P. Thidiazuron:A potent regulato r of in vitro plant morphogenesis[J]. In vitro cell dev biol-plant, 1998, 34(4):267-275. doi: 10.1007/BF02822732 URL
[18]	郭艳超, 张倩, 田传卫, 等. 香水月季类原球茎(PLBs)途径再生植株的研究[J]. 中国农业大学学报, 2008, 25(5):29-34.
[19]	瞿素萍, 王继华, 唐开学, 等. 5个切花月季品种的叶片离体培养和再生能力的基因型效应[J]. 西南大学学报(自然科学版), 2007, 51(12):108-112.
[20]	Caboni E, Tonelli M G. Effect of 1,2-benzisoxazole-3-acetic acid on adventitious shoot regeneration and in vitro rooting in apple[J]. Plant Cell Reports, 1999, 18(12):985-988. doi: 10.1007/s002990050695 URL
[21]	张绿萍. 刺梨花药和未受精胚珠离体培养[D]. 贵阳: 贵州大学, 2009.
[22]	李萍, 成仿云, 张颖星. 防褐剂对牡丹组培褐化发生、组培苗生长和增殖的作用[J]. 北京林业大学学报, 2008, 30(2):71-76.
[23]	陈爱萍, 冯维卫, 刘允康. 3种豆科牧草愈伤组织继代培养抗褐化的研究[J]. 中国农学通报, 2017, 33(22):138-144.
[24]	陈梦倩, 范李节, 王小德. 香港四照花外植体的抗褐化处理与诱导培养[J]. 浙江农林大学学报, 2018, 35(4):778-784.
[25]	蒲艳, 张鹏远, 王娟, 等. 滇牡丹成熟胚和叶柄愈伤组织的诱导及培养条件优化[J]. 分子植物育种, 2022, 20(1)1-18.
[26]	陈豆豆, 贺亮亮, 章鹏, 等. 无核品种‘中葡萄18号’胚挽救技术体系的建立及其杂交后代早期选择[J]. 果树学报, 2021, 38(12):2223-2235.

处理组合	浓度/(mg/L)	时间/h
M0	0	0
M1	50	8
M2	50	12
M3	100	8
M4	100	12

处理组合	浓度/(mg/L)	时间/h
M0	0	0
M1	50	8
M2	50	12
M3	100	8
M4	100	12

处理	基因型	诱导率/%	褐化率/%	生长状态
M0	贵农1号	87.30±0.17 c	10.34±0.05 a	致密颗粒状,黄绿色
M1		82.49±0.07 e	8.51±0.08 b	致密颗粒状,嫩绿色
M2		85.64±0.08 d	5.42±0.13 c	致密颗粒状,嫩绿色
M3		91.95±0.30 b	3.15±0.06 d	致密颗粒状,嫩绿色
M4		97.44±0.86 a	1.46±0.06 e	致密颗粒状,嫩绿色
M0	贵农2号	84.55±0.09 c	11.85±0.24 a	致密颗粒状,黄绿色
M1		80.27±0.06 e	9.02±0.02 b	致密颗粒状,嫩绿色
M2		83.33±0.26 d	7.66±0.11 c	致密颗粒状,嫩绿色
M3		85.92±0.24 b	4.50±0.12 d	致密颗粒状,嫩绿色
M4		89.37±0.20 a	2.41±0.06 e	致密颗粒状,嫩绿色
M0	贵农5号	84.55±0.24 d	9.55±0.26 a	致密颗粒状,黄绿色
M1		82.77±0.33 e	7.79±0.12 b	致密颗粒状,嫩绿色
M2		85.80±0.12 c	5.85±0.07 c	致密颗粒状,嫩绿色
M3		87.93±0.30 b	3.43±0.11 d	致密颗粒状,嫩绿色
M4		92.16±0.10 a	1.13±0.09 e	致密颗粒状,嫩绿色
M0	贵农7号	83.75±0.07 c	14.24±0.30 a	致密颗粒状,黄绿色
M1		79.54±0.48 d	10±0.17 b	致密颗粒状,嫩绿色
M2		84.61±0.13 c	7.56±0.05 c	致密颗粒状,嫩绿色
M3		90.56±0.11 b	5.47±0.10 d	致密颗粒状,嫩绿色
M4		94.00±0.82 a	3.53±0.08 e	致密颗粒状,嫩绿色

处理	基因型	诱导率/%	褐化率/%	生长状态
M0	贵农1号	87.30±0.17 c	10.34±0.05 a	致密颗粒状,黄绿色
M1		82.49±0.07 e	8.51±0.08 b	致密颗粒状,嫩绿色
M2		85.64±0.08 d	5.42±0.13 c	致密颗粒状,嫩绿色
M3		91.95±0.30 b	3.15±0.06 d	致密颗粒状,嫩绿色
M4		97.44±0.86 a	1.46±0.06 e	致密颗粒状,嫩绿色
M0	贵农2号	84.55±0.09 c	11.85±0.24 a	致密颗粒状,黄绿色
M1		80.27±0.06 e	9.02±0.02 b	致密颗粒状,嫩绿色
M2		83.33±0.26 d	7.66±0.11 c	致密颗粒状,嫩绿色
M3		85.92±0.24 b	4.50±0.12 d	致密颗粒状,嫩绿色
M4		89.37±0.20 a	2.41±0.06 e	致密颗粒状,嫩绿色
M0	贵农5号	84.55±0.24 d	9.55±0.26 a	致密颗粒状,黄绿色
M1		82.77±0.33 e	7.79±0.12 b	致密颗粒状,嫩绿色
M2		85.80±0.12 c	5.85±0.07 c	致密颗粒状,嫩绿色
M3		87.93±0.30 b	3.43±0.11 d	致密颗粒状,嫩绿色
M4		92.16±0.10 a	1.13±0.09 e	致密颗粒状,嫩绿色
M0	贵农7号	83.75±0.07 c	14.24±0.30 a	致密颗粒状,黄绿色
M1		79.54±0.48 d	10±0.17 b	致密颗粒状,嫩绿色
M2		84.61±0.13 c	7.56±0.05 c	致密颗粒状,嫩绿色
M3		90.56±0.11 b	5.47±0.10 d	致密颗粒状,嫩绿色
M4		94.00±0.82 a	3.53±0.08 e	致密颗粒状,嫩绿色

取样时间		贵农1号		贵农2号		贵农5号		贵农7号
季节	月份	诱导率/%	褐化率/%	诱导率/%	褐化率/%	诱导率/%	褐化率/%	诱导率/%	褐化率/%
春季	3	95.80±0.46 a	2.24±0.09 l	90.71±0.33 a	5.68±0.03 g	90.71±0.33 a	3.33±0.06 k	91.40±0.12 a	8.45±0.11 j
	4	94.72±0.07 ab	3.31±0.06 k	90.56±0.10 a	5.96±0.02 g	90.56±0.10 a	3.94±0.03 j	91.08±0.37 a	8.56±0.07 j
	5	95.47±0.41 a	3.61±0.12 j	90.15±0.14 a	6.58±0.15 g	90.15±0.14 a	4.96±0.25 i	90.02±0.08 b	9.17±0.10 i
夏季	6	71.71±0.33 gh	26.57±0.10 c	65.33±0.60 h	41.42±0.08 c	65.33±0.60 h	25.65±0.03 c	75.47±0.08 f	41.09±0.40 c
	7	72.22±1.42 g	28.69±0.14 b	66.91±0.22 g	45.54±0.12 b	66.91±0.22 g	29.82±0.04 a	74.10±0.08 h	42.37±0.11 b
	8	70.20±0.08 h	30.41±0.08 a	66.5±0.31 g	49.63±0.12 a	66.5±0.31 g	29.87±0.20 a	71.29±0.12 i	45.66±0.04 a
秋季	9	82.04±1.01 f	22.49±0.08 d	71.22±0.49 f	30.21±0.03 d	71.22±0.49 f	26.50±0.16 b	74.74±0.44 g	45.54±0.11 a
	10	85.36±0.59 e	20.16±0.01 e	72.64±0.26 e	27.51±0.11 d	72.64±0.26 e	24.54±0.10 d	75.31±0.12 fg	36.48±0.13 d
	11	86±0.18 e	15.74±0.06 f	73.5±0.68 d	23.41±0.14 e	73.5±0.68 d	20.43±0.04 e	76.44±0.04 e	33.26±0.02 e
冬季	12	87.74±0.23 d	12.33±0.01 g	76.14±0.37 c	19.60±0.31 f	76.14±0.37 c	14.25±0.08 f	77.37±0.07 d	30.36±0.11 f
	1	90.85±0.37 c	10.61±0.06 h	78.28±0.13 bc	17.59±0.13 f	78.28±0.13 bc	10.45±0.06 g	79.80±0.27 c	15.33±0.18 g
	2	93.10±0.11 b	8.78±0.10 i	78.27±0.57 b	8.48±0.21 g	78.27±0.57 b	7.27±0.05 h	79.85±0.29 c	10.36±0.06 h