培养基与光照对沼兰种子非共生萌发的影响

doi:10.3773/j.issn.1005-264x.2010.04.009

摘要/Abstract

摘要：

近年来由于灭绝性采挖和生态环境的恶化, 沼兰(Malaxis monophyllos)的自然资源遭到严重破坏, 加上沼兰种子具有萌发缓慢和萌发率低的特性, 已处于近危乃至易危的状态。为了繁殖和保护沼兰种质资源, 利用添加2%蔗糖、3%活性炭和0.8%琼脂的Knudson C (KC)培养基, 研究了萘乙酸(NAA)、6-苄基腺嘌呤(6-BA)、椰乳、香蕉泥、土豆泥和光照条件6种因素对沼兰种子非共生萌发的影响; 同时通过光学显微镜和扫描电镜观察了种子的外部形态。结果显示, 沼兰种皮细胞的平周壁方向结构平坦、没有纹络, 但是沿着细胞垂周壁方向细胞壁较厚。通过正交试验设计和冗余分析, 建立了沼兰种子非共生萌发的最佳方案, 即在含有4.5 mg·L^-1NAA、10 mg·L^-16-BA、8%香蕉泥和3%土豆泥的KC培养基上光照条件(光照强度为20 μmol·m^-2·s^-1, 光周期为12 h/12 h)下萌发。该方案的种子萌发率在90%以上。

关键词: 非共生萌发, 沼兰, 正交设计, 冗余分析, 种子形态

Abstract:

Aims Extensive collection, environmental deterioration, slow seed germination and low germination percentage have seriously degraded the natural resources of Malaxis monophyllos, resulting in a vulnerable situation for the species. Our objective is to establish an asymbiotic germination system of M. monophyllos seeds to protect and propagate the species.

Methods Seed morphology was investigated by optical microscope and scanning electron microscope. We used Knudson C medium supplemented with 2% sucrose, 3% activated charcoal and 0.8% agar as the basic medium to investigate the effects of α-naphthalene acetic acid (NAA), 6-benzylaminopurine (6-BA), coconut milk, meshed banana, meshed potato and light on seed germination via the orthogonal test.

Important findings Periclinal walls of testa cells were smooth and had no veins, but anticlinal walls were thick and had clear surface patterns. Through the orthogonal test and redundancy analysis, an optimal formula for asymbiotic seed germination was determined: the medium supplemented with 4.5 mg·L^-1 NAA, 10 mg·L^-1 6-BA, 8% meshed banana and 3% meshed potato in the light (12 h light/12 h dark at 20 μmol·m^-2·s^-1 of light intensity). Seed germination was > 90% with this formula.

Key words: asymbiotic germination, Malaxis monophyllos, orthogonal test, redundancy analysis, seed morphology

王瑞霞, 何明高, 宋松泉. 培养基与光照对沼兰种子非共生萌发的影响. 植物生态学报, 2010, 34(4): 438-443. DOI: 10.3773/j.issn.1005-264x.2010.04.009

WANG Rui-Xia, HE Ming-Gao, SONG Song-Quan. Effect of media and light on asymbiotic germination of Malaxis monophyllos seeds. Chinese Journal of Plant Ecology, 2010, 34(4): 438-443. DOI: 10.3773/j.issn.1005-264x.2010.04.009

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https://www.plant-ecology.com/CN/Y2010/V34/I4/438

图/表 6

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水平 Level	因素 Factor
水平 Level	A, NAA (mg·L^-1)	B, 6-BA (mg·L^-1)	C, 椰乳 Coconut milk (%, v/v)	D, 香蕉泥 Meshed banana (%, w/v)	E, 土豆泥 Meshed potato (%, w/v)
1	0	0	0	0	0
2	0.5	1	4	4	3
3	1.5	3	8	8	6
4	4.5	10	10	10	12

水平 Level	因素 Factor
水平 Level	A, NAA (mg·L^-1)	B, 6-BA (mg·L^-1)	C, 椰乳 Coconut milk (%, v/v)	D, 香蕉泥 Meshed banana (%, w/v)	E, 土豆泥 Meshed potato (%, w/v)
1	0	0	0	0	0
2	0.5	1	4	4	3
3	1.5	3	8	8	6
4	4.5	10	10	10	12

处理Treatment	A	B	C	D	E	光照 Light		黑暗 Darkness
处理Treatment	A	B	C	D	E	萌发率Germination (%)	圆球茎诱导率Protocorm (%)		萌发率 Germination (%)	圆球茎诱导率Protocorm (%)
1	1	1	1	1	1	10.75 ± 1.75^fg	0.00 ± 0.00^d	73.00 ± 2.00^b		7.61 ± 1.28^a
2	1	2	2	2	2	45.00 ± 3.50^c	17.97 ± 2.51^a	52.50 ± 3.50^d		1.34 ± 1.34^bc
3	1	3	3	3	3	40.00 ± 6.00^cd	3.64 ± 0.70b^cd	36.50 ± 1.50^e		0.00 ± 0.00^c
4	1	4	4	4	4	23.00 ± 4.00^ef	0.00 ± 0.00^d	74.00 ± 4.00^b		0.00 ± 0.0^0c
5	2	1	2	3	4	79.00 ± 3.00^b	7.89 ± 0.65^abc	76.50 ± 2.50^ab		0.00 ± 0.00^c
6	2	2	1	4	3	3.50 ± 2.50^g	0.00 ± 0.00^d	81.00 ± 5.50^ab		4.25 ± 0.75^ab
7	2	3	4	1	2	20.50 ± 3.50e^f	0.69 ± 0.69^cd	78.67 ± 1.67^ab		3.28 ± 0.04^bc
8	2	4	3	2	1	40.75 ± 3.75^cd	4.39 ± 1.02^abcd	2.50 ± 1.50^g		0.00 ± 0.00^c
9	3	1	3	4	2	10.75 ± 2.50^g	0.00 ± 0.00^d	2.00 ± 1.00^g		0.00 ± 0.00^c
10	3	2	4	3	1	19.67 ± 3.67ef	16.96 ± 4.46^ab	15.50 ± 4.00^f		0.00 ± 0.00^c
11	3	3	1	2	4	86.25 ± 4.75^ab	7.61 ± 3.93^abc	85.25 ± 2.25^ab		0.00 ± 0.00^c
12	3	4	2	1	3	28.25 ± 2.75^de	4.55 ± 1.33^abcd	2.50 ± 1.00^g		0.00 ± 0.00^c
13	4	1	4	2	3	18.50 ± 2.50^ef	0.00 ± 0.00^d	83.25 ± 3.75^ab		4.47 ± 0.70^ab
14	4	2	3	1	4	36.50 ± 1.00^cd	6.23 ± 2.23^abc	61.50 ± 3.50^c		2.74 ± 1.88^abc
15	4	3	2	4	1	30.25 ± 0.75^de	1.61 ± 1.61^cd	8.75 ± 1.25^fg		0.00 ± 0.00^c
16	4	4	1	3	2	93.25 ± 2.25^a	8.77 ± 5.11^abc	89.50 ± 2.00^a		5.02 ± 0.45^ab

处理Treatment	A	B	C	D	E	光照 Light		黑暗 Darkness
处理Treatment	A	B	C	D	E	萌发率Germination (%)	圆球茎诱导率Protocorm (%)		萌发率 Germination (%)	圆球茎诱导率Protocorm (%)
1	1	1	1	1	1	10.75 ± 1.75^fg	0.00 ± 0.00^d	73.00 ± 2.00^b		7.61 ± 1.28^a
2	1	2	2	2	2	45.00 ± 3.50^c	17.97 ± 2.51^a	52.50 ± 3.50^d		1.34 ± 1.34^bc
3	1	3	3	3	3	40.00 ± 6.00^cd	3.64 ± 0.70b^cd	36.50 ± 1.50^e		0.00 ± 0.00^c
4	1	4	4	4	4	23.00 ± 4.00^ef	0.00 ± 0.00^d	74.00 ± 4.00^b		0.00 ± 0.0^0c
5	2	1	2	3	4	79.00 ± 3.00^b	7.89 ± 0.65^abc	76.50 ± 2.50^ab		0.00 ± 0.00^c
6	2	2	1	4	3	3.50 ± 2.50^g	0.00 ± 0.00^d	81.00 ± 5.50^ab		4.25 ± 0.75^ab
7	2	3	4	1	2	20.50 ± 3.50e^f	0.69 ± 0.69^cd	78.67 ± 1.67^ab		3.28 ± 0.04^bc
8	2	4	3	2	1	40.75 ± 3.75^cd	4.39 ± 1.02^abcd	2.50 ± 1.50^g		0.00 ± 0.00^c
9	3	1	3	4	2	10.75 ± 2.50^g	0.00 ± 0.00^d	2.00 ± 1.00^g		0.00 ± 0.00^c
10	3	2	4	3	1	19.67 ± 3.67ef	16.96 ± 4.46^ab	15.50 ± 4.00^f		0.00 ± 0.00^c
11	3	3	1	2	4	86.25 ± 4.75^ab	7.61 ± 3.93^abc	85.25 ± 2.25^ab		0.00 ± 0.00^c
12	3	4	2	1	3	28.25 ± 2.75^de	4.55 ± 1.33^abcd	2.50 ± 1.00^g		0.00 ± 0.00^c
13	4	1	4	2	3	18.50 ± 2.50^ef	0.00 ± 0.00^d	83.25 ± 3.75^ab		4.47 ± 0.70^ab
14	4	2	3	1	4	36.50 ± 1.00^cd	6.23 ± 2.23^abc	61.50 ± 3.50^c		2.74 ± 1.88^abc
15	4	3	2	4	1	30.25 ± 0.75^de	1.61 ± 1.61^cd	8.75 ± 1.25^fg		0.00 ± 0.00^c
16	4	4	1	3	2	93.25 ± 2.25^a	8.77 ± 5.11^abc	89.50 ± 2.00^a		5.02 ± 0.45^ab