壳寡糖浸种对马铃薯微型薯芽生长和内源激素含量的影响

doi:10.3864/j.issn.0578-1752.2023.04.016

摘要/Abstract

摘要：

【目的】浸种处理是马铃薯生产中常用的促进块茎萌发的手段，然而浸种剂使用不当，会引起内源激素的失调，导致马铃薯高脚苗的发生。研究壳寡糖浸种对马铃薯微型薯芽生长及内源激素含量的影响，为壳寡糖作为马铃薯微型薯拌种剂的安全使用提供理论支持。【方法】以‘费乌瑞它’马铃薯微型薯为试材，分别用10、50和200 mg·L^-1（w/v）的壳寡糖溶液浸种处理，筛选促芽生长的最佳壳寡糖浓度，以50 mg·L^-1（w/v）浸种15 min为处理组（COS₅₀），蒸馏水浸种为阴性对照（Con），并选择易引起块茎芽细长且疯长的外源赤霉素3（15 mg·L^-1，w/v）浸种处理为阳性对照（GA₃）。测定各处理对微型薯发芽率、发芽势、芽长和芽直径的影响，并检测浸种处理前（BT）、萌发前期（BGS）、萌发期（GS）和伸长期（VG）顶芽内源激素含量的变化，用石蜡切片染色观察GS期顶芽细胞的形态，采用Pearson相关分析研究芽生长与内源激素含量的内在联系。【结果】50 mg·L^-1（w/v）壳寡糖浸种处理可有效促进微型薯块茎芽的萌发及生长，芽长较Con组显著提高了74.7%，芽直径较GA₃组显著提高了37.2%；并在浸种后第5天，块茎发芽率即达100%，分别较Con和GA₃组提高40.0%和17.0%，同时，COS₅₀浸种处理显著提高了微型薯的发芽势，在处理后第5天，发芽势分别较Con组和GA₃组提高了80.0%和28.6%，块茎表现为出芽早，出芽整齐。不同浸种处理微型薯顶芽分生组织的内源激素含量也存在显著差异，在VG期，COS₅₀处理组块茎芽分生组织的生长素（IAA）、细胞分裂素（CTK）和赤霉素（GA）的含量分别较Con组提高了11.4%、2.7%和2.5%，而脱落酸（ABA）的含量显著降低了3.2%（P<0.05）。GA₃处理组内源GA、CTK及IAA含量最高，COS₅₀组的IAA、CTK和GA含量分别低于GA₃组5.1%、8.4%和2.0%（P<0.05），ABA含量无显著降低。然而，COS₅₀处理显著提高了VG期微型薯顶芽分生组织中内源多胺（PA）的含量，分别较Con组和GA₃组提高了7.0%和2.8%。石蜡切片染色的结果显示，GA₃处理的顶芽分生组织细胞沿芽轴方向纵向伸长膨大形成细长扁平状的细胞，而COS₅₀组分生组织细胞膨大均匀，无细长扁平状，且与Con组的细胞形态相似。相关性分析显示，COS₅₀处理诱导的马铃薯微型薯芽的增长、增粗、发芽率及发芽势的提高与内源激素CTK、GA以及PA含量的升高呈显著正相关（P<0.05），且微型薯块茎发芽过程中PA的含量与ABA的含量表现极显著负相关（r=-0.785，P<0.01）。【结论】50 mg·L^-1（w/v）的壳寡糖浸种通过提高马铃薯微型薯芽组织中内源多胺、生长素、细胞分裂素以及赤霉素的含量提高了微型薯芽长、芽直径及发芽率，但壳寡糖处理引起的内源赤霉素含量显著低于易引起苗细长疯长的外源赤霉素浸种处理组，块茎芽分生组织的细胞均匀膨大，出芽整齐，芽长且粗壮，未表现芽细长、疯长现象。

关键词: 壳寡糖, 马铃薯微型薯, 浸种, 发芽, 内源激素

Abstract:

【Objective】 Seed soaking treatment is commonly used in potato production to promote tuber germination and bud growth. However, the improper use of seed soaking agents can trigger an imbalance of endogenous plant hormone levels, leading to the emergence of potato high-legged seedlings. In this study, the immersion effects on the growth and endogenous hormone content of potato minitubers were studied, so as to provide a theoretical basis for the safe use of chitooligosaccharide as a seed dressing agent for potato minituber. 【Method】The minitubers of Favorita were soaked with chitooligosaccharide solution at different concentrations of 10, 50, and 200 mg∙L^-1 (w/v), respectively, and the most effective concentration for promoting bud growth was screened. Seeds treated with 50 mg∙L^-1 (w/v) chitooligosaccharide for 15 min was the treatment group (COS₅₀), and the negative and positive control groups were treated with distilled water (Con) and 15 mg∙L^-1 gibberellin (GA₃), respectively. The germination rate, germination energy, bud length and bud diameter of tubers after seed soaking treatment were determined. The content of endogenous hormones in bud tissue was analyzed by ELISA before treatment (BT), before germination stage (BGS), germination stage (GS), and vigorous growth period (VG), respectively, and the morphology of terminal bud cells of GS was observed by paraffin section staining. Pearson correlation analysis was used to determine the intrinsic differences between shoot growth and endogenous hormone content. 【Result】Seed soaking with 50 mg∙L^-1 (w/v) chitooligosaccharide effectively promoted the germination and growth of tuber buds of the potato minitubers. The bud length and bud diameter of the treated samples were significantly higher than that of Con group (74.7%) and GA₃ group (37.2%). On the 5th day after soaking, the tuber germination rate reached 100%, which was 40.0% and 17.0% higher than the negative and the positive control, respectively. The germination energy of the minitubers was significantly enhanced. On the 5th day after treatment, the germination energy of minituber increased by 80.0% and 28.6% compared with Con and GA₃ groups, respectively. Different seed soaking treatments also showed significant differences in the endogenous hormone contents in the terminal bud meristem of minitubers. Compared with the negative control, COS₅₀ treatment elevated the levels of auxin, cytokinin, and gibberellin in the tubers during VG period, and the contents of these hormone increased by 11.4%, 2.7%, and 2.5%, respectively, while the content of abscisic acid decreased by 3.2% (P<0.05). The positive control treatment promoted the highest levels of endogenous gibberellin, cytokinin and auxin, and COS₅₀ treatment group had significantly lower levels of auxin, cytokinin and gibberellin than the positive control group by 5.1%, 8.4%, and 2.0%, respectively (P<0.05). No significant reduction was observed in the level of abscisic acid. However, the contents of endogenous polyamines in the apical bud meristems of the minitubers treated with COS₅₀ was 7.0% and 2.8% higher than those in the Con and GA₃ treatment groups, respectively. The results of paraffin section staining revealed that the morphological differences between the minitubers terminal bud cells was attributed to the different seed soaking treatments. The cells of the terminal bud meristem expanded resulting in elongated and flat shape in GA₃ treatment, while the cells of COS₅₀ treatment group showed a uniform shape, similar to the negative control. Furthermore, the sprouts growth, sprouts diameter, germination rate and energy of minitubers were positively correlated with the increase of cytokinin, gibberellin and polyamines contents induced by COS₅₀ treatment (P<0.05), while abscisic acid and ployamines content were significantly negatively correlated (r=-0.785, P<0.01). 【Conclusion】Seed soaking with chitooligosaccharide increased the contents of auxin, cytokinin, gibberellin, and polyamines in sprouts of minitubers. The treatment also increased the germination rate, sprouts length and sprouts diameter of potato minitubers. Uniform enlargement of meristematic cells in the sprouts of minitubers observation showed that COS₅₀ treatment did not affect the longitudinal elongation of apical bud meristematic cells during the expansion process, causing the bud to grow wildly. However, gibberellin content decreased significantly in seed soaked with 50 mg∙L^-1 (w/v) chitooligosaccharide compared with the treatment with exogenous gibberellin 3.

Key words: chitooligosaccharide, minitubers of potato, seed soaking, germination, endogenous hormones

冶楠, 朱艳, 赵元寿, 朱建宁, 门佳伟, 陈富, 孔德媛, 张卫兵, 宗元元, 李永才. 壳寡糖浸种对马铃薯微型薯芽生长和内源激素含量的影响[J]. 中国农业科学, 2023, 56(4): 788-800.

YE Nan, ZHU Yan, ZHAO YuanShou, ZHU JianNing, MEN JiaWei, CHEN Fu, KONG DeYuan, ZHANG WeiBing, ZONG YuanYuan, LI YongCai. Effects of Seed Soaking with Chitooligosaccharide on the Growth of Sprout and Endogenous Phytohormone Content in Potato Minitubers[J]. Scientia Agricultura Sinica, 2023, 56(4): 788-800.

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相关性 Correlation	芽长 Bud length	芽直径 Bud diameter	发芽势 Germination energy	发芽率 Germination rate	GA	ABA	IAA	PA	CTK
芽长 Bud length	1.00	0.729**	0.911**	0.827**	0.906**	0.046	-0.158	0.674*	0.935**
芽直径 Bud diameter		1.00	0.384*	0.651**	0.577*	0.369	-0.278	0.491*	0.702**
发芽势 Germination energy			1.00	0.764**	0.451*	-0.26	-.384*	0.420*	0.626**
发芽率 Germination rate				1.00	0.775**	0.13	-.524*	0.591*	0.823**
GA					1.00	-0.303	-0.229	0.571*	0.903**
ABA						1.00	-0.146	-0.785**	-0.097
IAA							1.00	-0.316	-0.239
PA								1.00	0.352
CTK									1.00