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林业科学 ›› 2016, Vol. 52 ›› Issue (2): 67-73.doi: 10.11707/j.1001-7488.20160208

• 论文与研究报告 • 上一篇    下一篇

大叶相思花粉离体萌发适宜条件及活力检测方法

詹妮, 黄烈健   

  1. 中国林业科学研究院热带林业研究所 广州 510520
  • 收稿日期:2015-04-03 修回日期:2015-05-29 出版日期:2016-02-25 发布日期:2016-03-25
  • 通讯作者: 黄烈健
  • 基金资助:
    国家"十二五"科技支撑计划项目(2012BAD01B0402)。

Conditions for in vitro Germination and Testing Method for Pollen Viability of Acacia auriculiformis

Zhan Ni, Huang Liejian   

  1. Research Institute of Tropical Forestry, CAF Guangzhou 510520
  • Received:2015-04-03 Revised:2015-05-29 Online:2016-02-25 Published:2016-03-25

摘要: [目的] 对大叶相思花粉离体萌发适宜条件以及花粉活力不同检测方法进行研究,为今后进一步开展人工控制授粉、选育优良的马大杂交新品种提供基础。[方法] 采回花穗的次日10:00之后,使用毛笔刷法收集大叶相思花粉;通过不同培养温度、不同蔗糖浓度和不同硼酸浓度处理后进行花粉离体萌发,筛选适宜大叶相思花粉离体萌发的条件;采用过氧化物酶法、I2-KI法以及花粉管离体萌发法对大叶相思花粉进行活力测定,探讨大叶相思花粉有效的活力测定方法。[结果] 大叶相思花粉在28℃下培养,其花粉平均萌发率为71.99%,花粉管平均长度为5.3 D(1 D=1倍的花粉粒长度),花粉管平均条数为6.2条,显著高于其他培养温度的处理。当培养基中含有200 g·L-1蔗糖时,大叶相思花粉萌发率为84.96%,其花粉管平均长度为5.8 D,花粉管平均条数为6.2条,显著高于含有其他蔗糖浓度的处理。在含有300 mg·L-1硼酸的培养基中培养大叶相思花粉,其平均萌发率为75.32%,花粉管平均长度可达4.8 D,平均花粉管条数为5.4条,显著高于在其他硼酸浓度下的处理。大叶相思花粉在培养温度30℃、200 g·L-1蔗糖、300 mg·L-1硼酸的培养基中萌发率为98.26%,花粉管长度均可达到10倍复合花粉粒的长度,最多可萌发出10条花粉管,显著高于其他处理。培养3 h的大叶相思花粉,萌发率为65.74%;培养6 h时,其萌发率为90.55%;培养24 h后,其萌发率趋于稳定,达到最高值98.26%。过氧化物酶法检测的花粉活力为99.67%,I2-KI法检测的花粉活力为99.00%,花粉管离体培养法检测的花粉活力为98.15%,3种方法检测的花粉活力无显著差异。[结论] 大叶相思花粉在不同的离体培养处理下,其萌发情况均不相同且差异显著,最佳离体萌发条件为200 g·L-1蔗糖+300 mg·L-1硼酸,培养温度为30℃,萌发率为98.26%。过氧化物酶法、I2-KI法以及花粉管离体培养法检测的花粉活力无显著差异。研究结果可为今后开展大叶相思的花粉收集、贮藏及活力测定等研究,以及开展人工控制授粉选育优良马大杂种相思新品种等研究提供重要依据。

关键词: 大叶相思, 花粉收集, 花粉离体萌发, 花粉活力测定

Abstract: [Objective] This study was aimed to studying the suitable conditions of in vitro pollen germination and the methods of pollen viability test for Acacia auriculiformis in order to provide a basis for hybrid breeding of A. auriculiformis and creating new breeding material by artificial pollination.[Methods] Pollen of A. auriculiformis was collected after 10:00 am on the next day of collection of flower clusters using the brush method. The pollen was germinated in vitro in conditions with different temperatures, the different concentrations of sucrose and boric acid, and suitable conditions for in vitro pollen germination of A. auriculiformis were chosen. Peroxidase method, I2-KI method and in vitro pollen germination method were used to detect the pollen viability of A. auriculiformis, and the effectiveness of these methods was assessed.[Results] When the pollen was cultured at 28℃, germination rate was 71.99%, the average length of pollen tubes was 5.3 D (1D=the length of pollen grain), the average number of pollen tubes was 6.2, which were significantly higher than those of the other culture temperatures. When the culture medium contained 200 g·L-1 sucrose, the pollen germination rate was 84.96%, the average length of pollen tubes was 5.8 D, the average number of pollen tubes was 6.2, which were significantly higher than those of the other sucrose concentrations. When the culture medium contained 300 mg·L-1 boric acid, the pollen germination rate was 75.32%, the average length of pollen tubes could reach 4.8 D, the average number of pollen tube was 5.4, which were significantly higher than those of the other boric acid concentrations. A. auriculiformis pollen in the culture medium at 30℃ culture temperature with 200 g·L-1 sucrose and 300 mg·L-1 boric acid had a germination rate of 98.26%, and the length of pollen tubes could be up to 10 times of the composite pollen length, and 10 pollen tubes could be produced at most, significantly higher than other treatments. When cultured for 3 h, the pollen germination rate was 65.74%, when cultured for 6 h, the pollen germination rate was 90.5%, and for 24 h the pollen germination rate tends to be stabilized, reaching the maximum rate of 98.26%. By peroxidase method, pollen viability was 99.67%, by I2-KI method it was 99%, and by in vitro pollen germination it was 98.15%. There were no significant differences among the viabilities detected by the three methods.[Conclusion] Among the different treatments of A. auriculiformis pollen, the pollen germinations were significantly different. The most suitable conditions for in vitro pollen germination of A. auriculiformis was 200 g·L-1 sucrose and 100 mg·L-1 boric acid and 30℃ culture temperature, in which the pollen germination rate was 98.26%. There were no significant difference among the pollen viabilities detected by peroxidase, I2-KI and in vitro pollen germination. The results provide a theoretical basis for collection, storage, and viability test of pollen of A. auriculiformis, and also an important foundation for controlled artificial pollination and breeding new hybrid varieties of Acacia in the future.

Key words: Acacia auriculiformis, pollen collection, in vitro pollen germination, pollen viability test

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