CN111685127A - 一种促使植物无性繁殖的诱导剂及其应用 - Google Patents

一种促使植物无性繁殖的诱导剂及其应用 Download PDF

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CN111685127A
CN111685127A CN202010563474.2A CN202010563474A CN111685127A CN 111685127 A CN111685127 A CN 111685127A CN 202010563474 A CN202010563474 A CN 202010563474A CN 111685127 A CN111685127 A CN 111685127A
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杨涛
赵疆
李鑫
杨晖
王治业
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Pingliang Qingsong Traditional Chinese Medicine Decoction Pieces Co ltd
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Abstract

本发明涉及生物技术领域，具体涉及一种促使植物无性繁殖的诱导剂及其应用。所述的诱导剂包括精氨酸1000份，萘乙酸20份，6‑苄基腺嘌呤20份，表油菜素内酯2份，硝普钠100‑300份，茉莉酸甲酯20份或水杨酸20份。所述的催芽剂包括赤霉素4+7 50份，吲哚乙酸10份，氟啶酮20‑40份。本发明所述的诱导剂，可以显著的提高母鳞茎上诱导产生的子鳞茎的个数；催芽剂显著提高了子鳞茎的发芽率，本专利所述的技术方案成功实现了人工诱导无性克隆繁殖，较传统的实生苗育苗方法，生产大田移栽用小种茎的时间缩短了1年，减少了培育时间，降低了种植的人力物力的投入。

Description

一种促使植物无性繁殖的诱导剂及其应用
技术领域
本发明涉及生物技术领域，具体涉及一种促使植物无性繁殖的诱导剂及其应用。
背景技术
甘肃贝母(F.przewalshii Maxim)是名贵药材川贝母的药源植物之一，主要分布于甘肃省岷县、武都、舟曲、榆中等地海拔2800米以上的高山灌丛和草丛中，冷凉荫湿是其分布区气候的主要特点。甘肃贝母生长缓慢、对环境要求苛刻、繁殖系数低、根腐病发生率高，导致人工种植产业的发展受到严重的制约。解决甘肃贝母的繁殖问题成为当务之急。自然条件下，贝母只能进行种子繁殖，种子存在形态和生理后熟，实生苗途径繁育小种茎需要2年时间，而且根腐病、立枯病等苗期病害严重，影响贝母幼苗的生长，同时，贝母幼苗易受到早春低温及倒春寒的影响，成苗率低，且农事操作费时、耗力、耗费人工，导致贝母的种植无法满足市场需求。
专利甘肃贝母试管小鳞茎培育的方法(CN201510393001.1)公开了一种通过组织培养的方法快速生产试管小鳞茎的技术，有效解决了原球茎诱导率低、试管小鳞茎培养周期长、步骤繁琐的问题，但生产成本高，而且需要专门的技术人员进行操作，对劳动者技能要求高；发明专利一种甘肃贝母鳞茎瓣繁殖方法(CN201410400906.2)公开了一种鳞茎分瓣，逐步变温贮藏的方法，具有操作简单、有效缩短甘肃贝母栽培周期的优点，但繁殖系数低。
针对上述技术问题，本发明提供了一种促使植物无性繁殖的诱导剂，在设施条件下育苗，喷施所述诱导剂，结合催芽剂处理，变温贮藏，实现了通过无性克隆途径生产种苗的技术，此技术具有育苗周期短、繁殖系数高、病害少、不受早春极端环境影响、农事操作简单的优点。
发明内容
本发明的目的之一是提供一种促使植物无性繁殖的诱导剂，所述的诱导剂包括精氨酸1000份，萘乙酸20份，6-苄基腺嘌呤20份，表油菜素内酯2份，硝普钠100-300份，茉莉酸甲酯20份或水杨酸20份。
优选地，所述的诱导剂中包括硝普钠200-300份。
优选地，所述的诱导剂中还包括羟丙基甲基纤维素0.5份。
优选地，所述的植物为药用植物。
优选地，所述的药用植物为贝母。
以精氨酸，萘乙酸，6-苄基腺嘌呤，表油菜素内酯，硝普钠，茉莉酸甲酯或水杨酸为有效成分的诱导剂在促使植物无性繁殖中的应用。
一种催芽剂，所述的催芽剂包括赤霉素4+7 50份，吲哚乙酸10份，氟啶酮20-40份。
优选地，所述的催芽剂包括赤霉素4+7 50份，吲哚乙酸10份，氟啶酮30-40份。
所述的催芽剂在提高植物发芽率中的应用。
一种通过无性克隆方式生产甘肃贝母种茎的方法，所述的方法包括如下步骤：
(1)鳞茎的采收与栽种：
于8月上中旬贝母枯黄倒苗后採挖，採收4年生以上开花甘肃贝母的鳞茎；流水冲洗，多菌灵浸泡，取出晾干表面水分，于8月下旬栽种在无土基质中，自然条件下越冬，贝母于次年2月下旬出苗；
(2)贝母的生长条件，日常管理与所述的诱导剂喷施
A.生长条件：最高温度不超过30℃，空气湿度70％以上；
B.日常管理：每20天浇灌一次水溶性肥料，浓度为500mg/L，氮磷钾比例为20:20:20；
C.喷施人工诱导剂：待贝母出苗后，叶面喷施诱导剂，每2周喷施1次，共喷施4次；
(3)变温贮藏
6月上旬贝母倒苗后，采收贝母鳞茎，肉眼可见在母鳞茎上开始有子鳞茎形成，将母鳞茎包埋在无土基质中，每天16±2℃处理12小时，8±2℃处理12小时，共处理2个月；
(4)子鳞茎催芽与低温贮藏
变温贮藏后，8月上旬挖出贝母，小心的剥下子鳞茎，室温条件下用催芽剂浸泡30分钟，取出后晾干表面水分，包埋于无土基质中，2±2℃贮藏1个月；
(5)子鳞茎播种与小种茎的收获
9月上旬，上述低温贮藏后的子鳞茎播种于无土基质中，生长条件与日常管理同步骤(2)；12月下旬地上部分逐渐枯萎后收获小鳞茎，用作大田栽培小种茎；
本发明的有益效果是：①本发明所述的诱导剂，能够促使贝母母鳞茎产生子鳞茎，每个母鳞茎最高能够产生18-24个子鳞茎，相较于对照组显著的提高了母鳞茎上诱导产生的子鳞茎的个数，为贝母的无性克隆提供基础；②本发明将贝母母鳞茎通过变温贮藏，促进了子鳞茎的生长，使得储藏结束母鳞茎上附着的子鳞茎直径达到3.5±1毫米；同时变温贮藏促进了母鳞茎营养物质向子鳞茎的转移，子鳞茎逐渐长大，为其心芽的发育贮备了足够的营养物质；③本发明所述的催芽剂显著提高了贝母子鳞茎的发芽率，发芽率高达87±2％，有效保证了9月上旬子鳞茎栽培后的出苗率；④本专利所述的技术方案成功实现了人工诱导贝母无性克隆繁殖，仅用1年育苗，便可使小鳞茎直径6±1毫米，用于大田播种，较传统的贝母育苗方法，本发明所述的技术方案减少了培育时间，降低了种植的人力物力的投入。
附图说明
图1母鳞茎上诱导出的子鳞茎
具体实施方式
下面的实施例可以进一步说明本发明，但不应理解为对本发明的限制，在不背离本发明精神和实质的情况下，对本发明方法、步骤或条件所作的修改或替换，均属于本发明的范围。
以下实验所使用的甘肃贝母采自榆中县(海拔2800米)，温室位于榆中县和平镇生物苑基地(海拔1800米)，试验中用到的激素、化学试剂均为化学纯。水溶性肥料购自德国摩尔化工集团中国有限公司。
所述的形态后熟指果实成熟时，种胚尚未发育成熟，没有形成完全的成熟胚形态，还需要一定的时间发育，才能形成完整的胚结构。
所述的生理后熟是指种子虽然表现出形态成熟特性，但因有抑制物质存在，不具备发芽能力，还需经过一定时间发育，才能形成发芽能力。
所述的无土基质由育苗草炭土、颗粒蛭石、稻壳炭体积比为3:2:1组成。
实施例一、一种通过无性克隆方式生产甘肃贝母种茎的方法
1.甘肃贝母的采收与温室栽种
2018年8月中旬，在榆中县马坡乡林缘和草灌丛中採挖野生开花甘肃贝母，用自来水洗净表面泥土，1000倍液多菌灵浸泡20分钟，晾干表面水分，播种于混合基质中，温室中培养，常规管理。2019年2月底开始出苗，3月上旬出苗率达到90％以上，苗高10±3厘米。
2.贝母的生长条件，日常管理与所述的诱导剂喷施
3月上旬苗高10±3厘米时开始人工诱导子鳞茎，温室条件控制、日常管理、诱导剂喷施方法如下：
①温室条件控制：最高温度不超过30℃，空气湿度70％以上。
②日常管理：每20天浇灌一次水溶性肥料，浓度为500mg/L，氮磷钾比例为20:20:20。
③喷施人工诱导剂：叶面喷施人工诱导剂，每2周喷施1次，共喷施4次，喷施时人工诱导剂中加入0.05％的羟丙基甲基纤维素。
实验组和对照组在相同的环境条件下，相同的管理措施，相同的喷施方法进行处理，实验组和对照组人工诱导剂配方分别如表1和表2，每处理贝母个数20个，重复3次。
表1各个实验组所喷施诱导剂组分(单位mg/L)：
Figure BDA0002547009260000041
表2各个对照组喷施诱导剂组分(单位mg/L)：
Figure BDA0002547009260000042
Figure BDA0002547009260000051
6月上旬贝母倒苗后，挖出贝母鳞茎，肉眼可见子鳞茎的发生，统计母鳞茎上产生子鳞茎的个数，结果如表3。
表3实验组和对照组的母鳞茎上诱导产生的子鳞茎数量统计
Figure BDA0002547009260000052
Figure BDA0002547009260000061
实验组和对照组的母鳞茎上诱导产生的子鳞茎数量统计如表3所示，相较于对照组，实验组的母鳞茎上附着的子鳞茎的个数显著提高。且实验组的母鳞茎上附着的子鳞茎的个数随着诱导剂的剂量增高而增多，子鳞茎变小，从对照组5和对照组6的实验结果可以得到，不使用诱导剂贝母不会产生子鳞茎，无法实现贝母的无性繁殖，单独使用硝普钠也不会产生子鳞茎，无法实现贝母的无性繁殖。
3.变温贮藏
将上述实验组附着有子鳞茎的贝母母鳞茎包埋在无土基质中，基质组成同上，放在培养箱中变温贮藏，相对湿度70±5％，每天16±2℃处理12小时，8±2℃处理12小时，共处理2个月。
如图1所示，变温贮藏后母鳞茎萎蔫发黄，子鳞茎直径3.5±1毫米。通过变温贮藏处理，促进了母鳞茎营养物质向子鳞茎的转移，子鳞茎逐渐长大，为其心芽的发育贮备了足够的营养物质。
4.子鳞茎催芽与低温贮藏
将上述变温贮藏后的母鳞茎取出，小心的剥下子鳞茎，室温条件下，使用催芽剂浸泡30分钟，晾干表面水分，包埋于无土基质中，2±2℃贮藏1个月。催芽处理方案见表4，每处理随机选取100个子鳞茎，重复3次。
表4不同组使用催芽剂的组成和浓度、发芽率、子鳞茎平均根数(单位mg/L)
Figure BDA0002547009260000062
Figure BDA0002547009260000071
注：大写字母代表极显著水平下(p<0.01)多重比较
子鳞茎催芽低温贮藏后，2019年9月上旬播种于无土基质中，上述温室条件下培养。播种后20天开始出苗。由表4可以看出，实验组的发芽率均高于78±3％，最高达到了87±2％，相较于对照组获得了显著提高，具有显著性差异，且实验组的子鳞茎的平均根数为3-4条，对照组每个子鳞茎平均1-2条根，实验组的平均根数高于对照组。
5.小种茎收获
子鳞茎播种后日常管理，每20天浇灌一次水溶性肥料，浓度为500mg/L，氮磷钾比例为20:20:20。设施条件最高温度不超过30℃，空气湿度70％以上。12月下旬地上部分逐渐枯萎后收获小种茎，直径6±1毫米，可用于大田播种。
实施例二、甘肃贝母实生苗传统方法育苗
2017年8月中旬，采收野生甘肃贝母种子，在甘肃贝母拟境栽培试验地(榆中县马坡乡，海拔2600米)搭设遮光度70％的遮阳网起畦育苗。每亩土地施用甘肃绿能农业科技股份有限公司生产的生物有机肥300公斤，畦宽1米，畦高10厘米，采收未完全发黄开裂的贝母果实，取出种子撒播于畦面，薄薄覆盖细土以种子不露出为度，然后覆盖麦草、松针等保墒，常规管理。2018年4月中旬出苗，7月中旬倒苗；2019年4月下旬出苗，7月底倒苗，种植2年小鳞茎直径4±0.5毫米。
综上所述，本发明所述的诱导剂，能够促使贝母母鳞茎产生子鳞茎，每个母鳞茎最高能够产生18-24个子鳞茎，相较于对照组显著的提高了母鳞茎上诱导产生的子鳞茎的个数，为贝母的无性克隆提供基础；本发明将贝母母鳞茎通过变温贮藏，促进了子鳞茎的生长，使得储藏结束母鳞茎上附着的子鳞茎直径达到3.5±1毫米；同时变温贮藏促进了母鳞茎营养物质向子鳞茎的转移，子鳞茎逐渐长大，为其心芽的发育贮备了足够的营养物质；本发明所述的催芽剂显著提高了贝母子鳞茎的发芽率，发芽率高达87±2％，有效保证了9月上旬子鳞茎栽培后的出苗率；本专利所述的技术方案成功实现了人工诱导贝母无性克隆繁殖，仅用1年育苗，便可使小鳞茎直径6±1毫米，用于大田播种，较传统的贝母育苗方法，本发明所述的技术方案减少了培育时间，降低了种植的人力物力的投入，而且小种茎病害很少、种茎更大、品质更优。

Claims (10)

1.一种促使植物无性繁殖的诱导剂，其特征在于，所述的诱导剂包括精氨酸1000份，萘乙酸20份，6-苄基腺嘌呤20份，表油菜素内酯2份，硝普钠100-300份，茉莉酸甲酯20份或水杨酸20份。
2.如权利要求1所述的诱导剂，其特征在于，所述的诱导剂中包括硝普钠200-300份。
3.如权利要求2所述的诱导剂，其特征在于，所述的诱导剂中还包括羟丙基甲基纤维素0.5份。
4.如权利要求3所述的诱导剂，其特征在于，所述的植物为药用植物。
5.如权利要求4所述的诱导剂，其特征在于，所述的药用植物为贝母。
6.以精氨酸，萘乙酸，6-苄基腺嘌呤，表油菜素内酯，硝普钠，茉莉酸甲酯或水杨酸为有效成分的诱导剂在促使植物无性繁殖中的应用。
7.一种催芽剂，其特征在于，所述的催芽剂包括赤霉素4+7 50份，吲哚乙酸10份，氟啶酮20-40份。
8.如权利要求7所述的催芽剂，其特征在于，所述的催芽剂包括赤霉素4+7 50份，吲哚乙酸10份，氟啶酮30-40份。
9.如权利要求7或8所述的催芽剂在提高植物发芽率中的应用。
10.一种通过无性克隆方式生产甘肃贝母种茎的方法，其特征在于，所述的方法包括如下步骤：
(1)鳞茎的采收与栽种：
于8月上中旬贝母枯黄倒苗后採挖，採收4年生以上开花甘肃贝母的鳞茎；流水冲洗，多菌灵浸泡，取出晾干表面水分，于8月下旬栽种在无土基质中，自然条件下越冬，贝母于次年2月下旬出苗；
(2)贝母的生长条件，日常管理与所述的诱导剂喷施
A.生长条件：最高温度不超过30℃，空气湿度70％以上；
B.日常管理：每20天浇灌一次水溶性肥料，浓度为500mg/L，氮磷钾比例为20:20:20；
C.喷施人工诱导剂：待贝母出苗后，叶面喷施诱导剂，每2周喷施1次，共喷施4次；
(3)变温贮藏
6月上旬贝母倒苗后，采收贝母鳞茎，肉眼可见在母鳞茎上开始有子鳞茎形成，将母鳞茎包埋在无土基质中，每天16±2℃处理12小时，8±2℃处理12小时，共处理2个月；
(4)子鳞茎催芽与低温贮藏
变温贮藏后，8月上旬挖出贝母，小心的剥下子鳞茎，室温条件下用催芽剂浸泡30分钟，取出后晾干表面水分，包埋于无土基质中，2±2℃贮藏1个月；
(5)子鳞茎播种与小种茎的收获
9月上旬，上述低温贮藏后的子鳞茎播种于无土基质中，生长条件与日常管理同步骤(2)；12月下旬地上部分逐渐枯萎后收获小鳞茎，用作大田栽培小种茎；
所述的诱导剂如权利要求1-5任一项所述，所述的催芽剂如权利要求7或8所述。
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