全 文 ：Research on Rapid Propagation and
Domestication of Wild Raspberry Plantlets
Weijun ZHU*, Senfu XU, Yitao CHEN
Taizhou Vocational College of Science & Technology, Taizhou 318020, China
Supported by Taizhou College Students’ Innovative Team Funding Project (class I).
*Corresponding author. E-mail: 714930911@qq.com
Received: August 28, 2015 Accepted: December 24, 2015A
Agricultural Science & Technology, 2016, 17(2): 298-300
Copyright訫 2016, Information Institute of HAAS. All rights reserved Molecular Biology and Tissue Culture
W ild raspberry is a woody plantof Rosaceae, commonlyknown as Shumei, Yemei,
Mumei and Wupaozi, and its ripe fruit
has the effects of tonifying liver and
kidney, securing essence and reducing
urination and improving eyesight by
using as a medicine or for eating[1]. The
propagation of raspberry generally
adopts cuttage and root-splitting
method in production, though these
methods are limited to the base num-
ber of fine varieties with a low propa-
gation coefficient, and susceptible to
virus accumulation, resulting in variety
degeneration which has a strong im-
pact on yield and quality[2-3]. In this stu-
dy, a series of experiments was con-
ducted by tissue culture method, aim-
ing at obtaining a large quantity of
high-quality plantlets in a short period
and providing a basis for large-scale
production[4-6].
Materials and Methods
Experimental materials
All raspberry materials were from
Linhai Huahan Fupenzi specialized
cooperatives, and the variety was
Rubus chingii Hu, the one-year-old
tender stem segments of which were
used as explants.
Experimental methods
Material pretreatment
The branches having leaves cut
were cleaned with soapy water and a
soft brush and then flushed with tap
water for 10 min, and the clean
branches were cut into segments,
each of which had one shoot.
Sterilization with ethanol
The cut stem segments were
transferred into sterile cups on a clean
bench; 70% ethanol was poured into
the sterile cups to immerse the mate-
rial, and poured out after shaking for 1
min; and the stem segments were
flushed with sterile water once.
Sterilization with HgCl2
Above material was immersed
with 0.1% HgCl2 and shaken for 6 -8
min, and then the HgCl2 solution was
poured out.
Cleaning with sterile water
A proper amount of sterile water
was poured into each sterile cup and
poured out after shaking for several
Abstract [Objective] This study was conducted to acquire a large quantity of wide
raspberry plantlets and provide a basis for large-scale production. [Method] Re-
search was performed on series tissue culture propagation of wide raspberry. [Re-
sult] Adventitious shoots were induced with stem segments, and the best induction
medium was MS +1.5 mg/L BA +0.3 mg/L NAA with an induction rate of 96%; and
the best differentiation medium was MS +2.0 mg/L KT +0.5 mg/L NAA, which ex-
hibited high root number and quality, with an average root number of 4.5 per plant.
The substratum for training plantlets was leaf mould 60%+perlite 30%+vermiculite
10%; and the temperature of 23-27 ℃ and relative air humidity of 80%-90% were
beneficial to survival of trained plantlets. [Conclusion] This study laid a foundation
for further large-area cultivation of Rubus chingii Hu.
Key words Wild raspberry; Tissue culture; Proliferation; Domestication
野生覆盆子种苗快繁与驯化试
验研究
朱伟俊*，徐森富，陈依桃 （台州科技职业学
院，浙江台州 318020）
摘 要 [目的]为短期内获得大量优质覆盆子
种苗及大规模生产提供依据。 [方法]对野生覆
盆子进行系列组培快繁研究。 [结果]利用茎段
能诱导出不定芽， 最佳诱导培养基为 MS +1.5
mg/L BA +0.3 mg/L NAA，诱导率可达 96％；最
佳分化培养基为 MS +2.0 mg/L KT +0.5 mg/L
NAA，芽生长健壮，增殖达到 4.2 倍；最佳生根
培养基为 l/2 MS +0.2 mg/L NAA，生根数量质
量好，平均每株生根 4.5 条。炼苗基质以 60％腐
叶土+30％珍珠岩+10％蛭石为佳 ； 炼苗温度
23～27 ℃，空气相对湿度 80％～90％有利于提高
炼苗成活率。 [结论]该试验为进一步进行掌叶
覆盆子的大面积栽培奠定了基础。
关键词 野生覆盆子；组织培养；增殖；驯化
基金项目 2014 年度台州市大学生科技创新
团队资助项目（I 类）。
作者简介 朱伟俊（1995-），男，浙江永嘉人 ，
专业：园艺技术，E-mail：714930911@qq.com。 *
通讯作者。
收稿日期 2015-08-28
修回日期 2015-12-24
DOI:10.16175/j.cnki.1009-4229.2016.02.013
Agricultural Science & Technology2016
Table 3 Effects of different culture media on rooting
Treatment Inoculation number Root number Rooting rate//%
M13 50 41 82
M14 50 45 90
M15 50 46 92
M16 50 44 88
Table 1 Effects of different culture media on induction
Treatment Inoculation number Number of induced shoots Shooting rate//%
M1 50 35 70
M2 50 46 92
M3 50 48 96
M4 50 42 84
Table 2 Effects of different culture media on differentiation
Treatment Inoculation number Reproduction number Reproduction multiple
M5 50 70 1.4
M6 50 80 1.6
M7 50 135 2.7
M8 50 165 3.3
M9 50 185 3.7
M10 50 210 4.2
M11 50 180 3.6
M12 50 150 3.0
times, and this operation was repeated
for 5 times. The water on the material
was sucked with aseptic water for later
use.
Culture medium and preparation
MS was used as a minimal medi-
um (containing sucrose 3% and agar
0.7%, pH 5.8), which was added with
different concentrations of hormones,
and the prepared culture media were
added into 150 ml glass bottles and
sterilized[7-8].
Results and Analysis
Pretreatment of explants
Plants with good characters were
selected, marked, and then subjected
to peripheral sterilization.
Experiment of young raspberry sh-
oot induction
Healthy raspberry plants were
collected on a sunny day, and subject-
ed to cleaning and sterilization treat-
ment, and the sterilized explants were
added into induction media for shoot-
induced cultivation.
The sterile tender stems with
shoots were cultured in the culture
media of MS plus combinations of I-BA
and different concentrations of 6-BA.
The experimental result showed that
the combination of MS+1.5 mg/L6-BA+
0.3 mg/LNAA was the most suitable
culture medium, in which axillary
shoots grew out 5 d after inoculation,
and grew to 2-2.5 cm after 25 d. The
axillary shoots were cut off for subcul-
ture (Fig. 1).
Experiment of proliferation of rasp-
berry plantlets
Induced explants were separated
from plants and inoculated into the
proliferation media. The result showed
that the combination of MS +2.0 mg/L
KT +0.5 mg/L NAA was the most suit-
able medium, in which shoots grew
healthy and strong, and the reproduc-
tion multiple reached 4.2 (Fig. 2).
Experiment of rooting of raspberry
plantlets
The proliferated young plantlets
were put into a rooting medium, in
which the plantlets grew into mature
plantlets satisfying the standard of
strong plantlet with vigorous roots.
After proliferation by subculture,
above best test-tube plantlets growing
vigorously and uniformly (with a length
of 1 cm, having 3 -4 small leaves)
were inoculated onto 1/2 MS added
with different concentrations of hor-
mone to perform rooting culture. White
root primordia generated from lower
cuts 5 d after inoculation, and the ten-
der roots were 2.5-4 cm in length after
20 d. During the experiment, the num-
ber of inoculated plantlets, formation
time of root primordia, number and
lengths of roots and number of rooted
plantlets were recorded. The experi-
ment result showed that the combina-
tion of l/2 MS+0.2 mg/L NAA was the
most suitable medium, which exhibited
a high rooting rate, better root quality
and an average root number of 4.5 per
plant.
Transplantation of raspberry plan-
tlets
Rooted intact healthy plants were
transplanted from tissue culture bottles
to perform domestication in green-
houses followed by training and plant-
ing. The substratum, leaf mould 60%+
perlite 30%+vermiculiten 10% was bet-
ter for training of the plantlets; and the
temperature of 23 -27 ℃ and relative
air humidity of 80%-90% were benefi-
cial to survival of trained plantlets. Dur-
ing the plantlet training, it was neces-
sary to control temperature and humid-
ity rigidly to control pests and dis-
eases.
When the rooted plantlets had de-
veloped root systems with plant
lengths up to 3-4 cm, they were trans-
ferred into greenhouses with the bot-
tles uncovered for 3 d, the culture
medium on the root systems was then
removed by cleaning, and the plantlets
were transplanted into nursery plates
with 72 cells. The used substratum
was adopted as turfy soil (or leaf
mould):perlite:vermiculite of 1:1:1. The
nursery plates were placed in a small
shade frame, which was removed 2
weeks later when the cells were full
with roots. The plantlets were trans-
planted into 10cruXl0cm nutrition pots,
and the survival rate was above 95%[9].
Discussion and Conclu-
sions
The nutritive elements needed for
preparation of the rooting medium
were only half of those required by pri-
mary culture and subculture, and the
used hormones were less as well [10].
The results of research on rapid prop-
agation of wild raspberry by tissue cul-
ture showed that: the best induction
medium was MS +1.5 mg/L BA +0.3
299
Agricultural Science & Technology 2016
Responsible editor: Yingzhi GUANG Responsible proofreader: Xiaoyan WU
[23] GUO YL, ROUX SJ. Partial Purification
and Characterization of a Ca2 +-De-
pendent Protein Kinase from the
Green Alga, Dunaliella salina[J]. Plant
Physiol, 1990, 94: 143-150.
[24] COWAN AK, ROSE PD. Abscisic acid
metabolism in salt-stressed cells of
Dunaliella salina [J]. Plant Physiol,
1991, 97(2): 798-803.
[25] YUASA T, MUTO S. Ca2+-dependem
protein kinase from the halotolerant
green alga Dunaliella tertiolecta: partial
purification and Ca2+-dependent asso-
ciation of the enzymeto the micro-
somes[J]. Arch Biochem Biophy, 1992,
296(1): 175-192.
Fig. 1 Induction of young raspberry shoots
Fig. 2 Experiment of proliferation of rasp-
berry plantlets
Responsible editor: Yingzhi GUANG Responsible proofreader: Xiaoyan WU
mg/L NAA; the best differentiation
medium was MS +2.0 mg/L KT +0.5
mg/L NAA; and the best rooting medi-
um was l/2 MS +0.2 mg/L NAA. Tissue
culture could provide a large quantity
of high-quality plantlets, laying a foun-
dation for further large-area cultivation
of Rubus chingii Hu.
References
[1] LIU WP (刘文萍 ).Rapid propagation
technique for virus-free plantlets of
Rosa chinensis (月季脱毒苗的快繁技
术 ) [J]. Chinese Agricultural Science
Bulletin (中国农学通报 ), 1997, 13(3):
59-60.
[2] WANG XR (王小蓉), TANG HR (汤浩
茹), DENG QX (邓群仙). Advancement
in research of genetic diversity of bram-
ble (Rubus L.) and its breeding in china
(中国树莓属植物多样性及品种选育研
究进展)[J]. Acta Horticulturae Sinica (园
艺学报), 2006(01): 190-196.
[3] XU E (徐娥), LI Y (李岩). Tissue Culture
and Rapid Propagation of Rubus cor-
chorifolius (树莓的组织培养及快速繁
殖 ) [J]. Chinese Wild Plant Resources
(中国野生植物资源), 2006(01): 64-65.
[4] YANG SH (杨少辉),JI J (季静),WANG G
(王 罡 ), et al. Tissue culture of Pha-
laenopsis and transplantation technique
by water culture (蝴蝶兰组织培养及水
培移栽技术)[J].天津大学学报(Journal of
Tianjin University), 2008(06): 709-713.
[5] PAN BR (潘彬荣), LUO TK (罗天宽),
ZHANG YX (张 永 鑫 ).Tissue culture
technique for Rubus chingii Hu (掌叶覆
盆子的组织培养技术 ) [J]. Journal of
Zhejiang Agricultural Sciences (浙江农
业科学), 2010(03): 508-510.
[6] CAO CY (曹春英). Plant tissue culture
(植物组织培养)[M]. Beijing: China Agri-
culture Press (北京: 中国农业出版社),
2006.
[7] LIU JP (刘进平).Tissue culture of tropi-
cal plants (热带植物组织培养)[M]. Bei-
jing: Science Press (北京 :科学出版社),
2006.
[8] LI S (李胜), LI W (李唯 ). Plant tissue
culture principle and technique (植物组
织培养原理与技术)[M]. Beijing: Chemi-
cal Industry Press (北京 :化学工业出版
社), 2008.
[9] HUANG Y (黄宇 ).Tissue culture and
rapid propagation technique of orchid
(兰花组织培养与快速繁殖技术 ) [M].
Beijing: China Agriculture Press (北京:
中国农业出版社), 2007.
[10] ZHENG CM, et al. Plant tissue culture
(植物组织培养)[M]. Hangzhou: Zhejiang
University Press (杭州: 浙江大学出版),
2011.
(Continued from page 293)
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
300