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花型花色单一、花期短、花叶均赏品种稀缺是木兰科Magnoliaceae植物杂交育种的主要问题[1]。目前有关含笑属Michelia杂交亲和性的研究较少,而了解物种间亲和性关系是培育新品种的关键。有研究认为:在含笑属远缘杂交过程中存在不亲和现象[2−3]。王亚玲等[4]则发现含笑属内种间杂交大多表现亲和,而紫花含笑M. crassipes×金叶含笑M. foveolata则不亲和,但未对不亲和原因进行探究。金叶含笑隶属木兰科含笑属,其花淡黄白色,花被片内扣形成独特花型,叶背密被红褐色柔毛,是木兰科中珍稀的花叶共赏树种,也是木兰科优良的杂交亲本材料。紫花含笑是含笑属珍稀的紫色花色植物,含笑M. figo的花被片边缘有紫色条纹。本研究通过金叶含笑与紫花含笑和含笑的杂交育种研究,以期将三者的优良性状相结合,培育出花叶共赏新品种,提高金叶含笑利用价值,丰富含笑属种质资源,并探究不亲和的原因,以期提高含笑属杂交成功率,为含笑属远缘杂交育种提供理论依据。
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如图1所示:授粉2 h时,仅有极少量的花粉附着在含笑柱头表面且花粉未萌发(图1A);4、6 h时,附着在含笑柱头表面的花粉数量逐渐增多,但同样未萌发(图1B和C);8 h时,有少量花粉开始萌发,花粉管长度为花粉粒直径的1.5~2.0倍(图1D);10 h时,萌发的花粉数量增加,花粉管长度为花粉粒直径的2.0~3.0倍,此时花粉管无汇聚现象(图1E);1 d时,花粉大量萌发,花粉管长度生长至花粉粒直径的3.0倍以上,花粉管出现了明显的汇聚现象(图1F);2 d时,花粉管生长至近花柱底部,即将进入子房(图1G);4 d时,花粉管进入子房,生长至子房1/3~1/2处(图1H);6 d时,有少数生长较快的花粉管进入胚珠(图1I);8 d时,大多数花粉管在此时已进入胚珠(图1J)。但同时也出现了花粉管生长异常的现象:授粉4 d时,发现花柱道细胞和间隙间堆积了大量胼胝质(图1K和L),阻碍了花粉管生长,这一现象也在4、8 d时出现;2 d时,花粉管应汇聚成束进入花柱,但有些花粉管在柱头表面生长杂乱无章(图1M)。在8 d时发现,花粉管在柱头表面萌发,但因花粉管内部堆积胼胝质而停止生长(图1N)。
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如图2所示:授粉2 h时,花粉未萌发(图2A);4 h时,柱头表面的花粉数量增多,有极少数花粉在此时开始萌发(图2B);6 h时,花粉萌发数量逐渐增多(图2C);在8 h时,大量花粉萌发,花粉管生长至花粉粒直径的3倍以上,此时花粉管存在汇聚的趋势但还没有开始汇聚(图2D);授粉后1 d,花粉管已汇聚成束,生长至柱头中部(图2E);2 d时,花粉管生长至花柱底部(图2F);4 d时,花粉管生长至子房约1/3处(图2G);6 d时,花粉管生长至子房1/2处(图2H);8 d时,大多数花粉管已进入胚珠(图2I)。该组合出现以下受精前障碍:授粉1 d时,出现花粉管生长杂乱且无汇聚现象。有的花粉萌发后,花粉管朝反方向生长(图2J);8 d时,发现有的花粉粒仍未萌发、萌发的花粉管在柱头表面彼此间交错缠绕生长,未进入柱头(图2K);花粉管在进入胚珠后,花粉管弯曲缠绕成团,阻碍受精(图2L);4 d时,花粉管内部和先端有胼胝质堆积而使花粉管膨大,阻碍其进一步生长(图2M)。
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如图3所示:授粉2 h时,无花粉黏附在柱头表面(图3A);4 h时,有少量花粉黏附在柱头表面,并开始萌发,花粉管长度为花粉粒直径的2~3倍(图3B);6 h时,萌发的花粉数量增加,花粉管生长至花粉粒直径的4倍以上,生长较快的花粉管已开始汇聚成束(图3C);8 h时,萌发的花粉数量显著增加,花粉管有明显汇聚成束的趋势(图3D);10 h时,花粉管汇聚成束(图3E);12 h时,汇聚成束的花粉管生长至近柱头下部(图3F);1 d时,成束的花粉管生长至子房上部1/3处(图3G);2 d时,成束的花粉管开始彼此分离,生长较快者已进入胚珠(图3H);4 d时,多数花粉管已进入胚珠(图3I);6 d时,花粉管逐渐向下生长,进入底部的胚珠(图3J)。6 d时,极个别的柱头表面仍存在大量未萌发的花粉,少数已萌发花粉的花粉管内存在胼胝质堆积(图3K)。
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如图4所示:授粉2 h后,仅有极少花粉黏附在柱头表面,无花粉萌发(图4A);4 h时,黏附的花粉数量增加,已有少量花粉萌发,花粉管生长至花粉粒直径的2~3倍,少数生长较快的花粉管生长至花粉粒直径的4倍以上(图4B);6 h时,黏附及萌发花粉数量大幅增加,花粉管生长至花粉粒直径的4倍以上(图4C);8 h时,花粉管有汇聚成束的趋势(图4D);10 h时,呈汇聚趋势的花粉管数量显著增加,汇聚趋势更加明显(图4E);12 h时,花粉管汇聚成束(图4F);1 d时,成束的花粉管生长至子房1/3~1/2处且花粉管开始彼此分离(图4G);2 d时,花粉管继续向下延伸(图4H);4 d时,多数花粉管已进入胚珠(图4I)。
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如表1和图5所示:金叶含笑与含笑正反交的坐果率均较高,分别为75.0%和78.8%。金叶含笑与紫花含笑正反交坐果率相差极大,正交的坐果率很高,达80.0%,而反交则极低,只有6.7%。金叶含笑×紫花含笑与金叶含笑×含笑的果实在坐果率、蓇葖膨大率与种子数上无显著差异,金叶含笑×含笑的种子较金叶含笑×紫花含笑略大而饱满。含笑×金叶含笑的坐果率显著高于紫花含笑×金叶含笑,蓇葖膨大率与种子数同样高于紫花含笑×金叶含笑。
表 1 金叶含笑在不同杂交组合中的结实情况
Table 1. Fruit and seed setting status of M. foveolata in different cross combinations
杂交组合 (♀×♂) 坐果率/% 结籽率/% 单个果序蓇葖
膨大率/%种子数/粒 单个蓇葖内
含种子数/粒形态异常
种子占比/%千粒重/g 种子萌发率/% 金叶含笑×含笑 75.0 a 50.3±13.4 a 90.5±8.8 a 300.7±79.2 a 4.8±1.1 a 9.6±5.8 a 25.1 90.6 金叶含笑×紫花含笑 80.0 a 36.6±9.6 b 87.6±7.1 a 287.5±47.9 a 4.4±0.6 a 16.1±9.2 a 21.8 78.1 含笑×金叶含笑 78.8 a 27.3±9.0 bc 49.4±15.7 b 24.1±9.7 b 1.1±0.2 b 0.0±0.0 b 107.7 87.5 紫花含笑×金叶含笑 6.7 b 18.4±1.4 c 27.3±4.6 c 13.0±1.4 b 1.1±0.0 b 0.0±0.0 b 113.3 92.9 说明:不同字母表示不同杂交组合间差异显著(P<0.05)。 -
于2021年和2022年9—12月采集各杂交组合的种子。含笑×金叶含笑与紫花含笑×金叶含笑的种子经低温沙藏处理后于翌年1月播种,种子在3月初至4月初开始陆续萌发。金叶含笑×含笑与金叶含笑×紫花含笑的种子属于薄皮种子,很容易因失水、氧化而失去活性,因此采用即采即播的方式,种子在翌年2月中旬陆续萌发。由表1所示:各杂交组合的种子出苗情况良好,萌发率较高,均在75.0%以上,其中最高的为紫花含笑×金叶含笑杂交组合的种子,萌发率高达92.9%。
Compatibility between Michelia foveolata and two Michelia species
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摘要:
目的 探明金叶含笑Michelia foveolata与含笑M. figo和紫花含笑M. crassipes的杂交亲和性。 方法 以金叶含笑与含笑和紫花含笑作正反交授粉,于授粉2 h、4 h、6 h、8 h、10 h、12 h、1 d、2 d、4 d、6 d、8 d后,通过荧光显微镜观察花粉管的生长情况。同时展开金叶含笑与含笑和紫花含笑的正反交杂交试验,统计结实情况。 结果 ①金叶含笑与含笑正反交的坐果率分别为75.0%和78.8%;②金叶含笑×紫花含笑坐果率(80.0%)显著高于紫花含笑×金叶含笑坐果率(6.7%);③含笑×金叶含笑花粉萌发较紫花含笑×金叶含笑晚2 h,花粉管在生长速度上无明显差异,均在8 d时进入胚珠;④以金叶含笑作母本的组合无明显杂交障碍,花粉管均在2~6 d内进入胚珠,种实发育良好;⑤含笑×金叶含笑存在花粉管在柱头表面杂乱生长、花粉管内部和先端有胼胝质堆积、花柱道细胞及间隙有大量胼胝质堆积等受精前障碍,虽不影响坐果,但对结籽有影响;⑥紫花含笑×金叶含笑存在受精前障碍和受精后障碍,受精前障碍包括花粉在柱头表面未萌发、花粉管在柱头表面盘旋生长未进入柱头、花粉管内部和先端堆积胼胝质而变得膨大,使花粉管生长受阻、花粉管在胚珠内弯曲缠绕成团,阻碍受精作用。但受精后障碍可能是影响落果的重要原因。 结论 金叶含笑与含笑正反交均表现亲和;金叶含笑×紫花含笑的亲和性程度高,紫花含笑×金叶含笑亲和性程度低。图5表1参23 Abstract:Objective This study is aimed to investigate the compatibility of Michelia foveolata with M. figo and M. crassipes. Method On the one hand, fluorescence microscope was used to observe the pollen tube growth after 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 1 d, 2 d, 4 d, 6 d and 8 d. On the other hand, the reciprocal cross test was carried out of M. foveolata with M. figo and M. crassipes respectively to calculate the fruit and seed setting status. Result (1) The fruit setting rates of the reciprocal cross of M. foveolata and M. figo were 75.0% and 78.8% respectively. (2) The fruit setting rate of M. foveolata × M. crassipes (80.0%) was significantly higher than that of M. crassipes × M. foveolata (6.7%). (3) The pollen germination of M. figo × M. foveolata was 2 h later than that of M. crasspies × M. foveolata and pollen tubes entered ovule on 8 d with no significant difference in growth rate. (4) There was no obvious hybridization barriers in the combination of M. foveolata as mother, and the pollen tube entered the ovule successively within 2 to 6 days with well developed fruit and seeds. (5) There were some pre-fertilization barriers, such as disarranged growth of pollen tube on stigma surface, callose accumulation in pollen tube, and callose accumulation in style channel cells and interstitial space and it was speculated that callose accumulation in style channel as the main cause of pre-fertilization barriers does not affect fruit setting, but has an effect on seed setting. (6) There were pre-fertilization and post-fertilization barriers in M. crassipes×M. foveolata. Pre-fertilization barriers include the failure of pollen to germinate on the stigma surface, the growth of pollen tubes in circles on the stigma surface, without entering the stigma, the expansion of pollen tubes due to the accumulation of callose inside and at the apex and the pollen tube becoming twisted and entangled within the ovule. However, post-fertilization barriers may be an important factor affecting fruit fall. Conclusion The reciprocal cross of M. foveolata and M. figo shows good compatibility with the compatibility degree of M. foveolata × M. crassipes being high, while the compatibility degree of M. crassipes×M. foveolata being low. [Ch. 5 fig. 1 tab. 23 ref.] -
Key words:
- Michelia foveolata /
- cross-compatibility /
- pollen tube /
- fruit setting rate /
- seed setting rate
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表 1 金叶含笑在不同杂交组合中的结实情况
Table 1. Fruit and seed setting status of M. foveolata in different cross combinations
杂交组合 (♀×♂) 坐果率/% 结籽率/% 单个果序蓇葖
膨大率/%种子数/粒 单个蓇葖内
含种子数/粒形态异常
种子占比/%千粒重/g 种子萌发率/% 金叶含笑×含笑 75.0 a 50.3±13.4 a 90.5±8.8 a 300.7±79.2 a 4.8±1.1 a 9.6±5.8 a 25.1 90.6 金叶含笑×紫花含笑 80.0 a 36.6±9.6 b 87.6±7.1 a 287.5±47.9 a 4.4±0.6 a 16.1±9.2 a 21.8 78.1 含笑×金叶含笑 78.8 a 27.3±9.0 bc 49.4±15.7 b 24.1±9.7 b 1.1±0.2 b 0.0±0.0 b 107.7 87.5 紫花含笑×金叶含笑 6.7 b 18.4±1.4 c 27.3±4.6 c 13.0±1.4 b 1.1±0.0 b 0.0±0.0 b 113.3 92.9 说明:不同字母表示不同杂交组合间差异显著(P<0.05)。 -
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