CN115226628A - 一种提高柑橘少核化育种效率的方法 - Google Patents
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Abstract
本发明提供了一种提高柑橘少核化育种效率的方法，通过加大辐射剂量，可明显增加辐射变异概率，可提高无核化育种效率，同时育种基数大且通过多种技术手段弥补辐射剂量过大导致的致死率，增加无核化育种基数；同时还通过多种手段来促进枝梢花芽分化，缩短育种筛选年限；并且对花芽分化后结果母枝进行体外促花，而后再进行花粉鉴定，缩短育种筛选年限。
Description
技术领域
本发明属于柑橘育种技术领域，具体涉及一种提高柑橘少核化育种效率的方法。
背景技术
国际上，1935年有人利用X射线进行柑橘诱变，1970年美国Hensz从热中子处理哈德逊葡萄柚中子的实生后代中选出无核、红肉的星红玉葡萄柚。我国柑橘辐射育种开始于20世纪60年代，得到了红江橙、雪柑等的无核或者少核突变体，黄建昌等人应用60Co-γ射线处理红江橙和沙田柚的接穗，试验结果表明：由于辐射诱发染色体联会消失，染色体变异，分裂不正常，导致花粉和胚胎败育，产生二倍体水平的无核突变。近年来陈善春等人进行辐射诱育柑橘无核品系的细胞遗传学研究，结果表明辐射诱发的染色体联会消失和结构变异引起的PMC减数分裂行为异常，致使花粉高度不育，从而表现出无核性状。但是由于辐射诱变变异概率很低，获得无籽品种存在不确定性与偶然性，因此国内育种机构多采用扩大辐射处理接穗量与嫁接单株来获得偶然的变异株，同时辐射接穗嫁接后采用自然生长挂果来鉴定是否有核或者少核，不但存在育种筛选周期长，还存在田间自然混交授粉无核品种有核化的误差。
发明内容
有鉴于此，本发明的目的在于提供一种提高柑橘少核化育种效率的方法，可解决辐射育种变异概率低、育种周期长、自然混交偏差等问题。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了一种提高柑橘少核化育种效率的方法，包括以下步骤：(1)对接穗进行辐照处理，得辐照接穗；所述辐照处理的浓度为1.21GY/min，处理时间为70min；
(2)将所述辐照接穗嫁接到砧木上；嫁接苗在花芽分化期，喷施6-BA促花芽分化；
(3)开花前进行催花，开花后进行花粉量鉴定，筛选花粉萌发率低于10％的嫁接苗进行后续培养。
优选的，步骤(1)所述接穗包含至少2个芽眼。
优选的，步骤(2)所述砧木的种植株行距为(0.5～1.0)m×(2～3)m。
优选的，步骤(2)所述嫁接包括多头嫁接，每株嫁接6～9个芽眼，且嫁接不成活时，重新补嫁接。
优选的，在步骤(2)所述嫁接成活后，还包括在秋梢老熟后将土壤含水量控制到12～18％，持续45天以上。
优选的，步骤(2)所述6-BA的工作液浓度为50～100ppm。
优选的，步骤(2)所述6-BA的工作液共喷施一次。
优选的，步骤(3)所述催花包括于27℃、光照量3000lx和湿度80％的环境中进行，所述催花的时间为一周。
有益效果：本发明提供了一种柑橘无核化快速育种方法，通过加大辐射剂量，可明显增加辐射变异概率，可提高无核化育种效率，同时育种基数大且通过多种技术手段弥补辐射剂量过大导致的致死率，增加无核化育种基数；同时还通过多种手段来促进枝梢花芽分化，缩短育种筛选年限；并且对花芽分化后结果母枝进行体外促花，而后再进行花粉鉴定，缩短育种筛选年限。
具体实施方式
本发明提供了一种提高柑橘少核化育种效率的方法，包括以下步骤：(1)对接穗进行辐照处理，得辐照接穗；所述辐照处理的浓度为1.21GY/min，处理时间为70min；
(2)将所述辐照接穗嫁接到砧木上；嫁接苗在花芽分化期，喷施6-BA促花芽分化；
(3)开花前进行催花，开花后进行花粉量鉴定，筛选花粉萌发率低于10％的嫁接苗进行后续培养。
本发明对接穗进行辐照处理，得辐照接穗；所述辐照处理的浓度为1.21GY/min，处理时间为70min。本发明对所述接穗的来源并没有特殊限定，优选至少包含2个芽眼，在进行后续嫁接时，可保证嫁接的成活率。本发明利用高于半数致死量的辐照浓度，可明显增加辐射变异概率，提高无核化育种效率。在本发明中，如采用半致死量进行辐照处理，辐射处理量为6000伦琴，辐射处理后接穗成活率在50％左右，枝梢变异率在60％左右，少核化变异概率大概在15％，但无核化变异概率不足1％；然而利用本申请的高浓度辐照，可将枝梢变异率提高至80％以上，少核化变异概率提高至30％，无核化变异概率2％。
得辐照接穗后，本发明将所述辐照接穗嫁接到砧木上；嫁接苗在花芽分化期，喷施6-BA促花芽分化。本发明所述砧木优选通过实生苗种植的方式获得，并且在种植时，采用密植的方式，获得大量砧木。本发明所述砧木的种植株行距优选为(0.5～1.0)m×(2～3)m，更优选为0.5m×2m。
本发明对所述嫁接的时机并没有特殊限定，根据本领域的常规时机进行嫁接，并且所述嫁接的方法优选包括多头嫁接，每株嫁接6～9个芽眼，且嫁接不成活时，重新补嫁接，所述补嫁接所用的接穗优选为经上述辐照并冷库保藏的接穗。
在所述嫁接成活后，本发明优选在秋梢老熟后将土壤含水量控制到12～18％，持续45天以上，更优选控制在15％。本发明所述控水有助于花芽分化，从而缩短育种年限。本发明优选还在花芽分化前，喷施6-BA，且所述6-BA的工作液浓度为50～100ppm，更优选为50ppm。本发明所述6-BA的工作液优选共喷施一次，有助于花芽分化，从而缩短育种年限。
本发明在开花前进行催花，开花后进行花粉量鉴定，筛选花粉萌发率低于10％的嫁接苗进行后续培养。本发明所述催花，优选包括于27℃、光照量3000lx和湿度80％的环境中进行，所述催花的时间为一周，更优选在人工气候箱中进行。在本发明中，花粉育性的高低是反映柑橘果实种子数的一个重要指标，辐射无核化育种主要原理也是使柑橘花粉母细胞在减数分裂期产生染色体易位、缺失及断裂等畸变，从而导致花粉败育形成少核或者无核，辐射处理后花粉败育比例显著高于对照，且随着剂量增大而降低。
下面结合实施例对本发明提供的一种提高柑橘少核化育种效率的方法进行详细的说明，但是不能把它们理解为对本发明保护范围的限定。
实施例1
1、采集十年生春香桔柚的枝条(春梢)进行辐射处理，浓度1.21GY/min，处理时间分别为30min、50min、70min，对照为无辐射处理。
2、采用行距2m×株距0.5m计划密植，亩种植基砧数在600株，砧木品种为三年生朱栾。
3、采用包含2个芽眼以上的接穗进行多头嫁接，单株嫁接芽眼量在6-9个之间，嫁接不成活后采用冷藏辐射接穗进行补嫁接。
表1不同辐射剂量处理后春香桔柚嫁接成活率
4、在秋梢老熟后将土壤含水量控制到15％左右，持续45天以上。
5、使用50ppm的6-BA进行喷叶处理促进花芽分化形成。
6、采用27℃，光照量3000lx，湿度80％的人工气候箱进行催花，一周后进行花粉萌发率鉴定，并对所结桔柚的种子数进行统计。
表2不同辐射剂量处理后春香桔柚花粉萌发率
表3不同辐射剂量处理后春香桔柚种子数
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (8)
1.一种提高柑橘少核化育种效率的方法，其特征在于，包括以下步骤：
(1)对接穗进行辐照处理，得辐照接穗；所述辐照处理的浓度为1.21GY/min，处理时间为70min；
(2)将所述辐照接穗嫁接到砧木上；嫁接苗在花芽分化期，喷施6-BA促花芽分化；
(3)开花前进行催花，开花后进行花粉量鉴定，筛选花粉萌发率低于10％的嫁接苗进行后续培养。
2.根据权利要求1所述方法，其特征在于，步骤(1)所述接穗包含至少2个芽眼。
3.根据权利要求1所述方法，其特征在于，步骤(2)所述砧木的种植株行距为(0.5～1.0)m×(2～3)m。
4.根据权利要求1所述方法，其特征在于，步骤(2)所述嫁接包括多头嫁接，每株嫁接6～9个芽眼，且嫁接不成活时，重新补嫁接。
5.根据权利要求1所述方法，其特征在于，在步骤(2)所述嫁接成活后，还包括在秋梢老熟后将土壤含水量控制到12～18％，持续45天以上。
6.根据权利要求1所述方法，其特征在于，步骤(2)所述6-BA的工作液浓度为50～100ppm。
7.根据权利要求1或6所述方法，其特征在于，步骤(2)所述6-BA的工作液共喷施一次。
8.根据权利要求2所述方法，其特征在于，步骤(3)所述催花包括于27℃、光照量3000lx和湿度80％的环境中进行，所述催花的时间为一周。
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| CN105684891A (zh) * | 2016-01-29 | 2016-06-22 | 江西农业大学 | 一种花粉辐射诱变创制无核柚的方法 |
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| CN105684891A (zh) * | 2016-01-29 | 2016-06-22 | 江西农业大学 | 一种花粉辐射诱变创制无核柚的方法 |
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| 周碧容等: "辐照诱变选育"无核金桔优株"", 《中国南方果树》 * |
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