第
35
卷第
7
期
2007
年
7
月
西北农林科技大学学报(自然科学版)
Journal
of
Northwest A
&.
F University(Nat.
Sci.
Ed.)
84
K
杨再生和遗传转化体系的优化
李科友,樊军锋,赵
忠,周永学,高建社
(西北农林科技大学林学院,陕西杨凌
712100)
Vo
l.
35
No.7
Ju
l.
2007
[摘 要]
为了优化
84
K
杨的再生和遗传转化体系,以
84
K
杨树叶片为外植体,采用正交试验研究了不同培养
基和激素配比对芽和芽苗生根的影响,并从预培养时间、浸染菌液浓度、浸染时间、共培养时间及添加
AS(
乙酷丁香
酣
)5
个方面,以卡那霉素抗性苗频率为衡量指标,研究了各因素对
84
K
杨遗传转化率的影响。结果表明
.84
K
杨叶
片最适的芽诱导培养基及激素配比为
MS+6-BA
0.5
mg/L+NAA
0.05
mg/L.
芽苗最适诱导生根培养基及激素配
比为
1/2
MS
十
NAA
0.05
mg/L+
IBA
0.05
mg/L;
优化筛选的最适转化体系为:农杆菌活化菌液用
MS
液体培养基
稀释
10
倍,浸染
5
min.
共培养
4
d
。采用上述优化的再生和遗传转化体系
.84
K
杨每片叶平均生芽数可达
20
个,芽
苗生根率可达
99%
.生根数为
6
条,叶盘转化率可达
35%
。
[关键词]
84
K
杨;再生系统;遗传转化;转化率
[中图分类号]
S792.119.04
[文献标识码]
A
[文章编号]
1671-9387(2007)07-0090-07
Prioritization of the system of regeneration and genetic
transformation of Populus
84
K
LI
Ke-you
,
FAN
Jun-feng
,
ZHAO
Zhong
,
ZHOU
Yong-Xue
,
GAO
Jian-she
(College
vf
Forestry.
N
ο
rthwestA
&.
F
University.Yangling.Shaanxi
712100.China)
Abstract:
To
prioritize
the
system
of
regeneration
and
genetic
transformation
of
Populus
84 K ,
influ-
ences
of
different
medium
and
proportion
of
hormone
on
bud
and
root
of
leaf-explant
were
studied
by
or-
thogonal
designing.
The
impact
of
five
aspects
(pre-culture
time
,
the
concentration
of
agro-infection
,
infect
time
,
co-culture
time
and
addition
of
AS
)
on
genetic
transformation
frequency
of
Populus
84 K
was
stud-
ied,
with
the
rate
of
kanamycin
resistance
seedling
as
the
measure
index.
The
results
demonstrated
that
the
optimal
differentiation
medium
and
proportion
of
hormone
for
leaf-explant
was
MS
十
BA
O.
5
mg/L+NAA
0.05
mg/L
,
and
the
optimal
rooting
medium
and
proportion
of
hormone
for
shoot
was
1/
2
MS+NAA
O.
05
mg/L+IBA
0.05
mg/
L.
The
optimal
prioritization
optimal
system
was
as
follows:to
use
MS
dilute
activa-
ted
agro-infection
for
10
times
,
to
infect
5
minutes
and
to
co-culture
for
4
days.
By
adopting
above
prioriti-
zation
system
of
regeneration
and
genetic
transformation
,
every
leaf
of
populus
84 K
could
be
induced
20
buds
on
average
,
rooting
rate
achieved
99 % ,
rooting
number
was
6,
and
transforming
rate
of
leaf
disc
was
35%.
Key
words:
Popul
us
84
K;
regeneration
system;
genetic
transformation;
transforming
rate
84
K
杨(银腺杨
,
Populus
alba
X
PO
ρ
ulus
glandulosa)
是南韩培育成功的一个著名白杨派内
[收稿日期]
2006-12-12
[基金项目]
科技部国家转基因植物研究与产业化专项
OY03-B-26
一
02)
[作者简介]
李科友
(]962-)
.男,陕西武功人,副教授,主要从事林木生物技术和植物化学研究。
[通讯作者]
樊军锋
(]962
一).男,陕西扶风人,副教授,主要从事林木遗传育种研究。