紫菜薹同源四倍体的诱导鉴定及特性研究

doi:10.16420/j.issn.0513-353x.2022-0686

摘要/Abstract

摘要：

为获得优质、抗病的同源四倍体紫菜薹（Brassica campestris var. purpurea Bailey）新材料，使用0.2 g · L^-1秋水仙素溶液点滴二倍体子叶顶端分生组织，经形态和解剖学、细胞学鉴定筛选得到同源四倍体植株。通过对二倍体、四倍体植株农艺性状和营养品质分析发现：四倍体植株的株高极显著增高；开展度、叶片宽、叶柄长显著增加；叶片数极显著降低。营养品质方面，可溶性蛋白、花青苷含量显著增高；可溶性糖和纤维素含量极显著降低；硝态氮含量显著降低。光合特性分析发现，四倍体植株的净光合速率、胞间CO₂浓度、气孔导度、蒸腾速率均高于二倍体，表明四倍体更能适应强光。抗病性分析发现，四倍体受灰霉菌侵染后，菌斑面积小，颜色淡，具有更强的抗病性。

关键词: 紫菜薹, 同源四倍体, 秋水仙素, 光合特性, 灰霉菌

Abstract:

In order to create new germplasm resource of tetraploid purple tsai-tai（Brassica campestris var. purpurea Bailey），0.2 g · L^-1 colchicine solution was used to drip the apical shoot of diploid. Then，autotetraploid plants were identified by morphology，anatomy and cytology methods. Through the analysis of agronomic characters，the plant height，spreading degree，leaf width and leaf stalk length of tetraploid plants increased significantly. However，the number of leaves of tetraploid significantly reduced. In nutritional quality，the content of soluble protein and anthocyanin in tetraploid plants increased significantly，while the content of soluble sugar，fibrin and nitric nitrogen in tetraploid plants significantly reduced compared with diploid. The analysis of photosynthetic characteristics showed that the net photosynthetic rate，interintracellular CO₂ concentration，stomatal conductance and transpiration rate of tetraploid plant were higher than that ofdiploid，indicating that tetraploid plants were more adaptable to strong light. The analysis of disease resistance showed that the tetraploid plants had stronger disease resistance.

Key words: Brassica campestris var. purpurea, autotetraploid, colchicine, photosynthetic characteristics, Botrytis cinerea

张晨, 李孟杰, 杨晓雪, 王美云, 肖栋, 王建军, 侯喜林, 胡君, 刘同坤. 紫菜薹同源四倍体的诱导鉴定及特性研究[J]. 园艺学报, 2023, 50(7): 1419-1428.

ZHANG Chen, LI Mengjie, YANG Xiaoxue, WANG Meiyun, XIAO Dong, WANG Jianjun, HOU Xilin, HU Jun, LIU Tongkun. The Creation and Study on Characteristics of New Material of Autotetraploid Purple Tsai-tai[J]. Acta Horticulturae Sinica, 2023, 50(7): 1419-1428.

图/表 9

表1 实时荧光定量PCR引物设计

Table 1 Primers for real-time PCR

引物名称 Primer name	引物序列（5′-3′） Primer sequence
Actin-Q	F：AGGCTACACGTTCGGACAAG
	R：TGGGGCACTAAACACAGTCA
Botrytis cinerea Actin-Q	F：AATGTTACCATACAAATCCTTACGGACA
	R：ACTCATATGTTGGAGATGAAGCGCAA

图1 紫菜薹二倍体（2x）与四倍体（4x）形态和解剖特征对比 A、B：植株；C：叶片；D：菜薹；E、F：花；G：荚果；H：种子；I、J：气孔；K、L：花粉粒。

Fig. 1 Comparison of morphological and anatomical differences between diploid and tetraploid purple tsai-tai A，B：Plant；C：Leaf；D：Tsai-tai；E，F：Floral organ；G：Silique；H：Seed；I，J：Stomata；K，L：Pollen.

图2 紫菜薹二倍体（2x）与四倍体（4x）根尖细胞染色体数鉴定

Fig. 2 Identification of chromosome number in root tip cells between diploid and tetraploid purple tsai-tai

图3 紫菜薹二倍体（2x）与四倍体（4x）流式细胞仪倍性鉴定结果

Fig. 3 Identification results between diploid and tetraploid purple tsai-tai by flow cytometry

表2 紫菜薹二倍体（2x）与四倍体（4x）植株主要农艺性状比较

Table 2 Comparison of main agronomic traits between diploid and tetraploid purple tsai-tai

材料 Material	株高/cm Plant height	叶片数 Number of leaves	单株质量/g Weight per plant	叶片长/cm Leaf length	叶片宽/cm Leaf width
2x	30.28 ± 0.50	18.20 ± 0.66	231.37 ± 30.79	16.58 ± 0.27	14.44 ± 0.17
4x	33.02 ± 0.54^**	14.40 ± 0.92^**	272.16 ± 29.98	17.02 ± 0.42	15.40 ± 0.31^*
材料 Material	叶柄长/cm Petiole length	叶柄宽/cm Petiole width	叶柄厚/mm Thickness of petiole	十叶厚/mm Ten leaves thickness	开展度/（cm×cm） Divergence of plant
2x	14.48 ± 0.74	1.10 ± 0.12	0.92 ± 0.04	4.10	（37.46 ± 3.08）×（38.12 ± 0.84）
4x	18.56 ± 0.20^*	1.36 ± 0.08	1.12 ± 0.08	4.83	（45.62 ± 1.33）×（38.24 ± 1.65）^*

表3 紫菜薹二倍体（2x）与四倍体（4x）营养品质比较

Table 3 Comparison of content of nutritional substance between diploid and tetraploid purple tsai-tai

营养指标 Nutritional index	材料Material
营养指标 Nutritional index	2x	4x
可溶性糖含量/（mg · g^-1）Soluble sugar content	23.46 ± 0.17	20.89 ± 0.09^**
可溶性蛋白含量/（mg · g^-1）Soluble protein content	26.90 ± 0.28	28.86 ± 0.36^*
纤维素含量/（μg · g^-1）Fibrin content	194.87 ± 1.19	111.28 ± 1.01^**
花青苷含量/（mg · g^-1）Anthocyanin content	0.13 ± 0.00	0.17 ± 0.01^*
叶绿素含量/（mg · g^-1）Chlorophyll content	9.74 ± 1.65	9.42 ± 0.80
硝态氮含量/（μg · g^-1）Nitric nitrogen content	13.79 ± 0.53	12.11 ± 0.25^*

图4 紫菜薹二倍体（2x）与四倍体（4x）植株光合特性比较

Fig. 4 Comparison of the photosynthetic characteristics between diploid and tetraploid purple tsai-tai

表4 紫菜薹二倍体（2x）与四倍体（4x）植株的光响应曲线参数的比较

Table 4 Comparison of the parameters of response curves between diploid and tetraploid purple tsai-tai

材料 Material	最大净光合速率/ （μmol · m^-2 · s^-1） P_n，max	表观量子效率 AQY	光饱和点/ （μmol · m^-2 · s^-1） LSP	光补偿点/ （μmol · m^-2 · s^-1） LCP	暗呼吸速率/ （μmol · m^-2 · s^-1） R_day
2x	32.06 ± 0.39	0.13 ± 0.14	257.51 ± 14.13	6.76 ± 0.76	0.68 ± 0.35
4x	47.54 ± 4.25^*	0.11 ± 0.03^*	487.48 ± 0.00^*	9.87 ± 1.71^*	1.12 ± 0.12

图5 紫菜薹二倍体（2x）与四倍体（4x）灰霉菌侵染菌斑（A）和灰霉菌Actin的表达量（B） * 表示在0.05水平差异显著。

Fig. 5 The Botrytis cinerea infection plaque of diploid and tetraploid purple tsai-tai（A）and expression of Botrytis cinerea Actin（B） * indicated significant differences at 0.05 level.

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