Methods for Reducing the Vitrification Rate and Promoting Rooting Formation of Dianthus plumarius

doi:10.7525/j.issn.1673-5102.2024.06.016

Abstract

Abstract:

The shoot tips of Dianthus plumarius were used as explants， and the optimal disinfection duration with HgCl? and the suitable proliferation medium were screened out. The effects of adding activated carbon（AC）， chlormequat（CCC）， polyvinylpyrrolidone（PVP）， and paclobutrazol（PP₃₃₃） on the vitrification aseptic seedlingsof D. plumarius， as well as the impact of different plant hormone ratios on rooting were investigated. The results indicated that the appropriate disinfection method for shoot tips of D. plumarius was 75% ethanol for 30 s followed by 0.1% HgCl₂ for 10 min， and the survival rate was 86.63%. The suitable proliferation medium was identified as MS supplemented with 2 mg?L^-1 6-benzylaminopurine（6-BA）， 0.05 mg?L^-1 Naphthaleneacetic acid （NAA）， 30 g?L^-1 sucrose， and 7 g?L^-1 agar， resulting in cluster buds proliferation rate of 90.28%， with an average bud height of 5.25 cm and a high proliferation coefficient of 12.31. In reducing the vitrification effect aseptic seedlingsof D. plumarius， the effect from high to low was CCC， AC， PP₃₃₃ and PVP. Adding 3 mg?L^-1 CCC in 1/2 MS medium was beneficial for overcoming the vitrification phenomenon and promoting plant growth. Adding 0.5 mg?L?1 NAA to the medium formula that is optimal for overcoming the vitrification rate is appropriate for the rooting of sterile seedlings of D. plumarius semperflorens. The rooting rate can reach 100%， with an average root length of 16.83 cm and an average number of 15.66 roots per individual plant. The findings provided theoretical and technical supports for the rapid propagation and industrial seedling production of D. plumarius.

Key words: Dianthus plumarius, sterilization, proliferation, vitrification, rooting

CLC Number:

Q949.745.8

Xuping TIAN, Jiali WANG, Ke QIAO, Huixin LIU, Kangjie YUE. Methods for Reducing the Vitrification Rate and Promoting Rooting Formation of Dianthus plumarius[J]. Bulletin of Botanical Research, 2024, 44(6): 954-962.

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References 26

1	韦莹，黄浩，黄宝优，等.植物生长抑制剂对短瓣石竹离体保存的影响［J］.中药材，2020，43（1）：24-27.
	WEI Y，HUANG H，HUANG B Y，et al，Effect of plant growth inhibitors on in vitro preservation of Dianthus brevivalvum ［J］.Journal of Chinese Medicinal Materials，2020，43（1）：24-27.
2	房立晶.香石竹的离体培养与试管开花研究［D］.重庆：西南大学，2008.
	FANG L J.Study on in vitro culture and in vitro flowering of Dianthus caryophyllus L.［D］.Chongqing：Southwest University，2008.
3	刘莹.‘娇媚三变’玫瑰快繁体系的建立及再生体系的初步研究［D］.泰安：山东农业大学，2022.
	LIU Y.Establishment of rapid propagation system and preliminary study on regeneration system of Rosa rugosa ‘Jiaomei Sanbian’［D］.Tai’an：Shandong Agricultural University，2022.
4	闫娜，陆艳辉，王春彦.石竹组织培养及试管苗诱导开花［J］.分子植物育种，2024，19（6）：1-8.
	YAN N， LU Y H， WANG C Y.Tissue culture and in vitro induced flowering of Dianthus chinensis ［J］.Molecular Plant Breeding，2024，19（6）：1-8.
5	BAYHAN N， YUCESAN B.Effects of sucrose and 6-benzylaminopurine concentrations on shoot regeneration and vitrification in Aronia melanocarpa：insights for plant tissue culture systems［EB/OL］.（2023-10-05）［2024-05-12］..
6	李佳慧，叶维雁，朱鹏锦，等.猫须草无菌短枝组织培养与快速繁殖体系的建立［J］.热带作物学报，2022， 43（10）：2063-2069.
	LI J H， YE W Y， ZHU P J，et al.Establishment of a sterile short shoot tissue culture and rapid propagation system for Clerodendranthus spicatus ［J］.Chinese Journal of Tropical Crops，2022，43（10）：2063-2069.
7	袁芳，李晶，任海平，等.藏药匙叶翼首草高频组织培养再生体系优化研究［J］.西北植物学报，2017，37（2）：379-386.
	YUAN F， LI J， REN H P，et al.Efficient in vitro plant regeneration system of Tibetan medicine Pterocephalus hookeri （C.B.Clarke） Höeck［J］.Acta Botanica Boreali-Occidentalia Sinica，2017，37（2）：379-386.
8	扁红英，苏世平，李毅，等.次氯酸钠对白刺开放式组培苗生理特性的影响［J］.草业科学，2022，39（2）：213-221.
	BIAN H Y， SU S P， LI Y，et al.Effect of sodium hypochlorite on physiological characteristics of Nitraria tangutorum open tissue culture seedlings［J］.Pratacultural Science，2022，39（2）：213-221.
9	KHARRAZI M， NEMATI H， TEHRANIFAR A，et al. In vitro culture of carnation （Dianthus caryophyllus L.） focusing on the problem of vitrification［J］.Journal of Biological and Environmental Sciences，2011，5（13）：1-6.
10	侯少霞，赵永秀，李佳雯，等.生长调节剂对香石竹离体快繁的影响［J］.分子植物育种，2018，16（3）：938-942.
	HOU S X， ZHAO Y X， LI J W，et al.Effects of different growth regulators on the rapid propagation in vitro of Dianthus caryophyllus ［J］.Molecular Plant Breeding，2018，16（3）：938-942.
11	KARAMI O.Induction of embryogenic callus and plant regeneration in carnation （Dianthus caryophyllus L.）［J］. Journal of Biological Sciences，2008，8（4）：68-72.
12	童雨嘉.满天星快速繁殖与农杆菌转化体系的建立［D］.武汉：华中农业大学，2019.
	TONG Y J.Establishment of rapid propagation and agrobacterium-mediated transformation in Gypsophila paniculata ［D］.Wuhan：Huazhong Agricultural University，2019.
13	韦莹，黄浩，董青松，等.短瓣石竹茎段诱导及植株再生研究［J］.中药材，2015，38（9）：1810-1812.
	WEI Y， HUANG H， DONG Q S，et al.Study on stem segment induction and plant regeneration of Brachystemma calycinum D.Don［J］.Journal of Chinese Medicinal Materials，2015，38（9）：1810-1812.
14	李莹.瞿麦组织培养与快繁技术的研究［D］.长春：吉林农业大学，2012.
	LI Y.Tissue culture and rapid propagation of Dianthus superbus L.［D］.Changchun：Jilin Agricultural University，2012.
15	高弘扬.石竹和拟南芥试管苗玻璃化机制及控制研究［D］.大连：大连理工大学，2018.
	GAO H Y.Mechanism and control of hyperhydricity in Dianthus chinensis L.and Arabidopsis thaliana［D］.Dalian：Dalian University of Technology，2018.
16	饶雪琴，张曙光.番木瓜组织培养中玻璃化苗的发生与预防［J］.植物生理学报，2005，41（3）：331.
	RAO X Q， ZAHNG S G.Occurrence and prevention of vitrification seedlings in Carica papaya tissue culture［J］.Plant Physiology Journal，2005，41（3）：331.
17	姜琳.五星花组织培养及试管开花研究［D］.北京：北京林业大学，2018.
	JIANG L.Study on tissue culture and in vitro flowering of Pentas lanceolata ［D］.Beijing：Beijing Forestry University，2018.
18	吕侃俐.长筒花、长寿花试管开花诱导及影响因素研究［D］.北京：北京林业大学，2017.
	LV K L.Study on influence factors of in vitro flowering induction of Achimenes spp.and Kalanchoe blossfeldi- ana［D］.Beijing：Beijing Forestry University，2018.
19	毛红俊.牡丹组织培养技术及玻璃化防止措施研究［D］.洛阳：河南科技大学，2011.
	MAO H J.Research on tissue culture technology and vitrification prevention measure for Paeonia suffruticosa ［D］.Luoyang：Henan University of Science and Technology，2011.
20	韩淑兰，王慧梅，张丹.降低黄芩再生苗玻璃化率及其生根的研究［J］.植物研究，2016，36（1）：90-96.
	HAN S L， WANG H M， ZAHNG D.Decreasing the hyperhydricity and rooting of Scutellariae baicalensis Georgi［J］.Bulletin of Botanical Research，2016，36（1）：90-96.
21	ZHANG J M， HUANG B， LU X X，et al.Cryopreservation of in vitro-grown shoot tips of Chinese medicinal plant Atractylodes macrocephala Koidz.using a droplet-vitrification method［J］.CryoLetters，2015，36（3）：195-204.
22	GHASHEEM NAL， STĂNICĂ F， PETICILĂ A G，et al. In vitro influence of cultivars and different culture media on vitrification and darkness on peach （Prunus persica L.Batsch） shoots multiplication［J］.Scientific Bulletin：Series F：Biotechnologies，2022，26（1）：59-64.
23	ROYCHOUDHRY S， KEPINSKI S.Auxin in root development［J］.Cold Spring Harbor Perspectives in Biology，2022，14（4）：309-333.
24	卢静洁，包希吉乐，张梦迪，等.生长调节剂对香石竹（Dianthus caryophyllus）组培苗生根的影响［J］.分子植物育种，2021，19（16）：5465-5469.
	LU J J， BAOXI J L， ZAHNG M D，et al.Effect of growth regulators on rooting of carnation（Dianthus caryophyllus） tissue culture seedlings［J］.Molecular Plant Breeding，2021，19（16）：5465-5469.
25	兰伟，蔡健，冯守平.常夏石竹的组培与快速繁殖［J］.林业科技开发，2006，20（2）：55-57.
	LAN W， CAI J， FENG S P.Mass propagation of Dianthus plumarius L. by tissue culture［J］.China Forestry Science and Technology，2006，20（2）：55-57.
26	黄冬华，周群，陶秀花，等.矮生香石竹的组织培养和快速繁殖［J］.中国农学通报，2007，23（9）：103-106.
	HUANG D H， ZHOU Q， TAO X H，et al.Tissue culture and rapid propagation of the short carnation［J］.Chinese Agricultural Science Bulletin，2007，23（9）：103-106.

处理号 No.	时间 Time/min	污染率 Contamination rate/%	成活率 Survival rate/%	褐化率 Browning rate/%
1	7	53.73±11.28^a	43.58±6.71^c	2.69±0.51^c
2	8	39.33±8.86^ab	49.98±2.31^c	10.69±2.66^b
3	9	18.19±4.79^b	64.28±3.96^b	17.53±4.21^ab
4	10	0^c	86.63±4.57^a	20.30±6.55^a
5	11	0^c	80.75±5.39^ab	23.37±2.69^a

编号 No.	活性炭质量浓度 AC/（g⋅L^-1）	玻璃化率 Vitrification rate/%	株高 Height/cm	叶片数量 Number of leaves	苗生长情况 Seedling growth status
A	0	86.13±15.41^a	5.89±1.26^c	6.33±2.15^c	叶片多干枯黄叶，玻璃化严重，植株细弱
B	0.5	61.67±14.71^b	6.69±1.45^bc	8.40±2.20^bc	叶片干枯黄叶较多，玻璃化严重，植株细弱
C	1.0	25.36±7.26^c	11.43±2.19^a	14.53±2.78^a	叶片明显增大多嫩绿，玻璃化极少，植株健壮
D	2.0	27.67±9.96^c	8.55±2.17^b	10.73±2.82^b	叶片增大多嫩绿，黄叶较少，玻璃化少，植株健壮
E	3.0	33.47±12.95^bc	7.48±1.61^b	9.20±2.17^b	叶片增大嫩绿，黄叶较少，玻璃化较少，植株细弱
F	4.0	40.33±12.47^bc	6.98±1.40^b	8.96±2.46^bc	叶片卷曲，黄叶多，玻璃化较重，植株细弱

编号 No.	矮壮素质量浓度 CCC/（mg⋅L^-1）	玻璃化率 Vitrification rate/%	株高 Height/cm	叶片数量 Number of leaves	苗生长情况 Seedling growth status
A	0	86.13±15.41^a	5.89±1.26^c	6.33±2.15^c	叶片多干枯黄叶，玻璃化严重，植株细弱
B	0.5	53.59±11.23^b	6.37±1.81^bc	8.52±1.33^bc	叶片嫩绿，玻璃化较重，植株较弱
C	1.0	17.85±5.22^c	7.96±1.25^b	10.81±2.69^b	叶片嫩绿，玻璃化较少，植株健壮
D	2.0	15.82±4.79^c	8.69±1.65^ab	12.66±1.89^a	叶片嫩绿，玻璃化较少，植株健壮
E	3.0	1.97±0.32^d	9.87±2.03^a	12.97±1.54^a	叶片嫩绿，玻璃化极少，植株健壮，茎增粗
F	4.0	14.69±3.37^c	9.06±1.77^a	12.55±1.75^a	叶片嫩绿，玻璃化极少，植株健壮

编号 No.	聚乙烯吡咯烷酮质量浓度 PVP/（mg⋅L^-1）	玻璃化率 Vitrification rate/%	株高 Height/cm	叶片数量 Number of leaves	苗生长情况 Seedling growth status
A	0	86.13±15.41^a	5.89±1.26^c	6.33±2.15^bc	叶片多黄叶干枯，玻璃化严重，植株细弱
B	0.5	81.73±10.63^a	6.69±0.94^bc	6.19±1.52^bc	叶片多黄叶干枯，玻璃化严重，植株细弱
C	1.0	72.69±11.31^b	8.52±1.19^a	8.21±1.71^b	叶片黄叶较多，玻璃化严重，植株细弱
D	2.0	54.97±10.08^c	8.94±2.01^a	10.35±2.03^a	叶片较绿，玻璃化较重，植株细弱
E	3.0	60.41±11.06^b	7.32±1.22^ab	8.06±1.22^b	叶片较绿，玻璃化较重，植株细弱
F	4.0	67.89±9.85^bc	5.51±0.87^c	6.07±0.82^bc	叶片嫩绿，玻璃化较重，植株细弱

编号 No.	多效唑质量浓度 PP₃₃₃/（mg⋅L^-1）	玻璃化率 Vitrification rate/%	株高 Height/cm	叶片数量 Number of leaves	苗生长情况 Seedling growth status
A	0	86.13±15.41^a	5.89±1.26^c	6.33±2.15^c	叶片多黄叶干枯，玻璃化严重，植株细弱
B	0.5	76.85±9.83^ab	6.22±0.81^b	6.98±1.46^bc	叶片多黄叶干枯，玻璃化严重，植株细弱
C	1.0	46.77±8.92^b	7.47±1.13^a	8.60±2.02^b	叶片嫩绿，玻璃化较重，植株细弱
D	2.0	31.82±9.88^bc	7.14±1.39^a	11.57±1.49^a	叶片嫩绿，玻璃化较少，植株健壮
E	3.0	26.64±9.77^c	6.13±1.34^ab	9.43±2.05^ab	叶片嫩绿，玻璃化少，植株健壮
F	4.0	58.43±10.19^b	5.21±1.01^bc	6.43±0.92^bc	叶片卷曲，玻璃化较重，植株细弱