德克萨斯组铃铛铁线莲胚胎挽救及促进种子萌发技术研究

孙瑞琦; 余伟军; 曾黎辉

doi:10.19303/j.issn.1008-0384.2019.09.007

德克萨斯组铃铛铁线莲胚胎挽救及促进种子萌发技术研究

doi: 10.19303/j.issn.1008-0384.2019.09.007

1.
福建农林大学，福建　福州　350002
2.
福建林业职业技术学院，福建　南平　353000

详细信息

作者简介:
孙瑞琦（1994−），男，硕士，研究方向：花卉与景观园艺（E-mail：308601900@qq.com）

通讯作者:
曾黎辉（1973−），女，博士，教授，研究方向：花卉基因工程、花色调控（E-mail：125865014@qq.com）

中图分类号: S 567
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出版历程
- 收稿日期: 2019-04-03
- 修回日期: 2019-08-09
- 刊出日期: 2019-09-01

Preliminary Studies on Embryo Rescue and Seed Germination Promotion of Clematis Texensis

1.
Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Fujian Forestry Vocational & Technical College, Nanping, Fujian　353000, China

摘要

摘要: 目的研究不同植物生长调节剂配比处理对德克萨斯组铃铛铁线莲打破休眠及胚胎挽救的影响，解决德克萨斯铃铛铁线莲萌发时间过长，高温胁迫引起种子发育中途败育的问题，为提高育种效率，缩短育种周期提供理论依据与指导。方法以德克萨斯组铃铛铁线莲‘王梦’为材料，设计12种不同浓度6-BA、NAA的正交试验筛选铃铛铁线莲胚胎挽救最适植物生长调节剂浓度，讨论椰乳作为培养添加物对胚胎挽救的促进作用；比较低温、变温及不同浓度GA、6-BA对打破铃铛铁线莲种子休眠的影响。结果将授粉后45 d的杂交铃铛铁线莲种子离体培养，MS基本培养基添加0.5 mg·L⁻¹ 6-BA和0.05 mg·L⁻¹ NAA可以使铁线莲胚胎正常发育成熟的成功率达到47%，发芽率为54%；在此基础上培养基中再添加10%的椰乳，种子发育成熟率和发芽率均显著提高到62%。低温处理和光培养25℃/暗培养15℃的温度可以促进铃铛铁线莲种子发芽，配合50 mg·L⁻¹ GA或30 mg·L⁻¹ 6-BA处理打破休眠更加显著。结论铁线莲胚可以通过离体挽救解决高温带来的败育问题；植物生长调节剂处理可以显著打破种子休眠，促进萌发。
- 铃铛铁线莲 /
- 杂交 /
- 打破休眠 /
- 胚胎挽救
Abstract: Objective To seek means for improvement on crossbreeding the appealing, long flowering, widely adaptable bell clematis by overcoming the long dormancy and high embryo abortion in seed germination. Method Seeds of Dream King clematis (Clematis texensis) collected 45 d after pollination were cultured in vitro on various media to optimize the conditions for embryo rescue and dormancy breaking. An orthogonal experiment of 12 treatments consisting of additions of 6-BA and/or NAA in various concentrations as well as exposures to temperatures and daylight was conducted to observe the resulting seed germination and seedling survival. Result The addition of 0.5 mg·L⁻¹ 6-BA and 0.05 mg·L⁻¹ NAA to the MS basic medium improved the seed germination rate to 54%, with 47% of the seedlings reached maturation. When 10% of coconut milk was added to the medium, both seed germination and seedling survival rate were significantly raised to reach 62%. Exposure of the seeds to low temperature and day 25℃/night 15℃ promoted the germination. And, the presence of either 50 mg·L⁻¹ GA or 30 mg·L⁻¹ 6-BA in the medium facilitated breaking the seed dormancy. Conclusion Embryos of clematis could be rescued in vitro to mitigate premature abortion caused by high temperature. The growth hormone addition could significantly shorten dormancy and promote germination of the bell clematis seeds.
- bell clematis /
- hybrid /
- seed dormancy and seed gernimation /
- embryo rescue

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图 1 未完成双受精过程的胚

Figure 1. Embryos failed to complete double fertilization

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图 2 发育至45天杂交铁线莲种子

Figure 2. Seeds of clematis hybrid developed after 45 d

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图 3 胚胎挽救正常发育种子

Figure 3. Embryo rescue of normally developed seeds

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图 4 胚胎挽救败育（异常膨大）种子

Figure 4. Embryo rescue of aborted seeds (Abnormal enlargement)

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图 5 胚胎挽救败育（干瘪）种子

Figure 5. Embryo rescue of aborted seeds (wizened)

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图 6 胚胎挽救成功种子、败育种子对比

Figure 6. Embryo-rescued and aborted seeds

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图 7 胚胎挽救成功后播种出苗

Figure 7. Embryo-rescued seeds after sowing and germination

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表 1 不同消毒时间对铁线莲种子的影响

Table 1. Effect of disinfection time on clematis seeds

消毒时间/min Sterilization time	污染率/% Pollution rate	褐变率/% Browning rate
8	70.00±0.20 a	8.33±0.03 c
10	30.00±0.10 b	22.66±0.08 bc
12	16.66±0.08 bc	33.33±0.08 b
15	0±0 c	63.33±0.19 a
注：同列数据后不同小写字母表示差异达显著水平（P＜0.05），下同。 Note: different lowercase letters after the data in the same column mean the difference reaches a significant level (P＜0.05)，the same as below.

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表 2 不同植物生长调节剂组合对铁线莲胚胎发育的影响

Table 2. Effect of hormone additions on clematis embryonic development

编号 Number	配方 Formula	发育率 Growth rate/%	发芽率 Germination rate/%
1	0.1 mg·L⁻¹ 6-BA+0.02 mg·L⁻¹ NAA	20.00±0.10 e	33.33
2	0.1 mg·L⁻¹ 6-BA+0.05 mg·L⁻¹ NAA	28.33±0.16 de	17.64
3	0.1 mg·L⁻¹ 6-BA+0.1 mg·L⁻¹ NAA	26.66±0.08 de	43.75
4	0.3 mg·L⁻¹ 6-BA+0.02 mg·L⁻¹ NAA	25.00±0.10 e	60.00
5	0.3 mg·L⁻¹ 6-BA+0.05 mg·L⁻¹ NAA	33.33±0.08 ced	60.00
6	0.3 mg·L⁻¹ 6-BA+0.1 mg·L⁻¹ NAA	30.00±0.09 de	44.44
7	0.5 mg·L⁻¹6-BA+0.02 mg·L⁻¹NAA	28.33±0.08 de	64.71
8	0.5 mg·L⁻¹6-BA+0.05mg·L⁻¹NAA	46.66±0.13 abc	53.57
9	0.5 mg·L⁻¹ 6-BA+0.1 mg·L⁻¹ NAA	41.60±0.13 bcd	52.00
10	0.5 mg·L⁻¹ 6-BA+0.02 mg·L⁻¹ NAA+10%椰乳（Coconut milk）	50±0.1 ab	50.00
11	0.5 mg·L⁻¹ 6-BA+0.05 mg·L⁻¹ NAA+10%椰乳（Coconut milk）	61.66±0.06 a	62.16
12	0.5 mg·L⁻¹ 6-BA+0.1 mg·L⁻¹ NAA+10%椰乳（Coconut milk）	48.33±0.1 abc	62.06

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表 3 低温、温差处理对铃铛铁线莲‘王梦’发芽率的影响

Table 3. Effect of low temperature and temperature differentiation on germination of clematis seeds

处理方法 Processing method	40 d 发芽率 Germination rate at 40 days/%
CK	0
低温 Low temperature	15.00±0.05 a
昼夜温差 Temperature difference	23.33±0.06 a

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表 4 植物生长调节剂处理对铃铛铁线莲‘王梦’发芽率的影响

Table 4. Effect of phytohormones on germination of clematis seeds

处理方法 Processing method	40 d 发芽率 Germination rate at 40 days/%
CK	36.66±0.06 cd
5 mg·L⁻¹ GA	35.00±0.10 c
20 mg·L⁻¹ GA	38.33±0.08 cd
30 mg·L⁻¹ GA	30.00±0.13 d
50 mg·L⁻¹ GA	76.66±0.14 a
80 mg·L⁻¹ GA	28.33±0.08 d
5 mg·L⁻¹ 6-BA	21.66±0.16 d
20 mg·L⁻¹ 6-BA	68.33±0.06 ab
30 mg·L⁻¹ 6-BA	81.66±0.06 a
50 mg·L⁻¹ 6-BA	61.66±0.13 ab
80 mg·L⁻¹ 6-BA	55.00±0.20 bc

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