百合种球繁育二次抽薹的淀粉代谢研究

杨成龙; 方少忠; 林智敏; 郭文杰; 蔡宣梅; 张洁

doi:10.19303/j.issn.1008-0384.2019.10.006

百合种球繁育二次抽薹的淀粉代谢研究

doi: 10.19303/j.issn.1008-0384.2019.10.006

福建省农业科学院生物技术研究所，福建福州 350003

基金项目: 福建省科技计划公益类专项（2018R1019-3）

详细信息

作者简介:
杨成龙（1990−），男，硕士，研究实习员，主要从事园艺作物栽培生理研究（E-mail：28613609@qq.com）

通讯作者:
张洁（1983−），女，硕士，助理研究员，主要从事园艺作物栽培生理研究（E-mail：zj5820212@163.com）

中图分类号: S 682
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出版历程
- 收稿日期: 2019-09-02
- 修回日期: 2019-10-03
- 刊出日期: 2019-10-01

Starch Metabolism of Repeated-bolting Lily Bulbs

Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350003, China

摘要

摘要: 目的研究百合种球反季节繁育二次抽薹过程中的淀粉代谢机制，为精确种球反季节繁育的收储期，促进国产种球繁育提供理论依据。方法通过测量百合抽薹前后不同发育时期，鳞茎中的淀粉、还原糖含量，使用扫描电镜观察鳞片细胞中的淀粉颗粒形态内容变化，定量分析不同发育时期种球的淀粉酶基因的表达情况。结果百合鳞茎二次抽薹过程中，淀粉和还原糖含量在萌动期出现显著性变化，淀粉含量下降4.98%，还原糖含量增加3.23%。扫描电镜（SEM）观察结果表明：百合鳞片抽薹过程中淀粉粒有A、B、C（大、中、小）3 种类型。在膨大期和成熟期包含A型和B型淀粉粒；萌动期C型淀粉粒出现，A型淀粉粒出现异形、凹陷或是拉伸，长短轴比（L/S）达最大1.77，淀粉颗粒被逐渐拆分；抽薹期A型淀粉粒消失，B、C型颗粒表面出现大量泡状突起，小颗粒可见以“出芽”的方式长出。不同发育时期淀粉酶基因的实时荧光定量多聚核苷酸链式反应（qRT-PCR）结果显示：淀粉分解酶基因AMY在不同发育时期均有表达，在膨大期和成熟期表达量较低，在萌动期和抽薹期高度表达；淀粉合成酶相关基因SSS、SBE在不同发育时期均有表达，在膨大期和成熟期表达量较高，在萌动期和抽薹期表达量较低；淀粉分解酶和合成酶相关基因在种球不同发育时期表达情况与淀粉含量的变化情况相符。结论百合种球反季节繁育过程中，二次抽薹导致种球大量的淀粉被消耗，将影响商品球质量以及鲜切花的品质，为消除此不利因素，繁育过程中应当在种球进入成熟期后及时进行收储。
- 百合 /
- 种球 /
- 淀粉代谢 /
- 抽薹
Abstract: Objective Starch metabolism at time of repeated bolting was studied to determine the optimum time for bulb-harvesting in propagation of out-of-season lilies. Method Contents of starch and reducing sugars as well as images of starch granules under scanning electron microscope of the lily bulbs at various developmental stages were examined. Expressions of amylase-related genes of the bulbs were obtained by qRT-PCR. Result The starch and reducing sugars in the bulbs underwent significant changes at germination stage, as the starch content decreased by 4.98% and reducing sugars increased by 3.23%. The SEM images of the starch granules were of A-, B- and C-types, or, large, medium and small in sizes. They were mainly A- and B-types in the expanding and mature bulbs. Upon germinating, the granules began to separate from one another with the appearances of C-type and deformed A-type that showed depressions or were stretched to an L/S ratio as high as 1.77. At bolting, the A-type granules disappeared while the B- and C-types appeared with bubbles and tiny " bud-like” growth on their surfaces. The qRT-PCR analysis indicated that the AMY gene in the bulbs expressed at a high-level during germination and bolting stages, while SSS and SBE at expanding and mature stages, and that the variations paralleled the contents of starch and reducing sugars. Conclusion A large amount of starch was consumed during repeated bolting of the bulbs. Thus, to reduce the ill-effects on the quality of the bulbs and lily flowers, bulb-harvesting at the mature stage was recommended.
- lily /
- bulb /
- starch matabolism /
- bolting

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图 1 百合鳞茎淀粉含量变化

注：不同小写字母表示差异显著（P＜0.05），相同字母者表示差异不显著（P＞0.05）。

Figure 1. Changes on starch content in lily bulbs

Note: Data with different lowercase letters indicate significant differences at P＜0.05; and, same letters indicate no significant difference at P＞0.05. The same for Fig. 2, 4, 5, and 6.

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图 2 百合鳞茎还原糖含量变化

Figure 2. Changes on reducing sugar content in lily bulbs

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图 3 百合鳞茎抽薹过程中淀粉粒的SEM图像

注：1~4. 膨大期（1）、成熟期（2）、萌动期（6）、抽薹期（4）百合中层鳞片淀粉粒的整体状态（×150）；5. 膨大期鳞片淀粉粒与絮状物共存（×500）；6. 成熟期鳞片淀粉粒（×500）；7. 萌动期鳞片淀粉粒出现形态变异（×500）；8. 抽薹期鳞片淀粉颗粒表面及腔体内壁出现大量泡状突起（×500）；9. 成熟期鳞片单个淀粉粒（×2 000）；10. 萌动期鳞片单个淀粉粒（×1 500）； 11. 抽薹期鳞片单个淀粉粒（×4 500）；12. 抽薹期鳞片出现淀粉粒呈撕裂状（×500）。

Figure 3. SEM images of starch granules in bolting lily buds

Note: 1-4. Whole starch granules in mid-layer scales of lily bulb at expanding stage (1), mature stage (2), germination stage (3), and bolting stage (4) (×150); 5. Starch granules and flocs at expanding stage (×500); 6. Starch granules at mature stage (×500); 7. Morphological variations of starch granules at germination stage (×500); 8. Surface and vesicular protuberances on surface and inner wall of starch granules at bolting stage (×500); 9. Individual starch granule at mature stage (×2 000); 10. Individual starch granule at germination stage (×1 500); 11. Individual starch granule at bolting stage (×4 500); 12. Torn starch granules at bolting stage (×500).

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图 4 不同发育时期百合鳞茎淀粉分解酶基因AMY相对表达水平

Figure 4. Relative expressions of AMY gene in lily bulbs at developmental stages

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图 5 不同发育时期百合鳞茎淀粉合成酶基因SSS相对表达水平

Figure 5. Relative expressions of SSS gene in lily bulbs at developmental stages

下载: 全尺寸图片幻灯片

图 6 不同发育时期百合鳞茎淀粉合成酶基因SBE 相对表达水平

Figure 6. Relative expressions of SBE gene in lily bulbs at developmental stages

下载: 全尺寸图片幻灯片

表 1 引物序列及内参基因信息

Table 1. Genes and primer sets used in qRT-PCR analysis

基因名称 Gene name	引物序列（5′–3′） Primer sequence	长度 Length/bp
AMY	F: GGGATATTCAGCAGAAATGGCGAAG	130
AMY	R: GCGGTTGGCATCTTGATTGTAGAGT	130
SSS	F: GTCAACGATTGGCACGCAAGTCT	171
SSS	R: ACCATACCACTCAGAGGGCAAGC	171
SBE	F: AGCAGCGGAGCGTCGTTAATCT	213
SBE	R: CCTGAGAGTCTTCAGCAGCCACT	213
ACTIN	F: GTCTGCGACAATGGTACTGGAATG	149
ACTIN	R: GCCTCATCACCAACATAAGCATCTT	149
注：F：正向引物；R：反向引物。 Note: F, forward primer; R, reverse primer.

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表 2 百合抽薹过程中淀粉粒特性比较

Table 2. Characteristics of starch granules of bolting lily bulbs

时期 Period	淀粉粒类型 Type of starch granule	长轴 Long axis(L)		短轴 Short axis(S)		L/S
时期 Period	淀粉粒类型 Type of starch granule	范围 Min-Max/μm	平均 Mean/μm	范围 Min-Max/μm	平均 Mean/μm	范围 Min-Max/μm	平均 Mean/μm
膨大期 Expanding stage	A	52.81-58.94	55.87 c	36.14-37.42	36.78 b	1.46-1.58	1.52 b
膨大期 Expanding stage	B	31.15-39.58	35.03 a	17.86-33.32	24.22 ab	1.16-1.88	1.45 a
成熟期 Mature stage	A	52.28-63.40	58.34 b	36.33-49.42	43.00 a	1.24-1.63	1.37 c
成熟期 Mature stage	B	21.08-48.68	35.60 a	14.01-36.52	25.37 a	1.21-2.25	1.50 a
萌动期 Germination stage	A	53.03-68.90	61.48 a	24.79-43.30	35.98 b	1.47-2.42	1.77 a
	B	22.00-49.66	29.49 b	17.28-35.92	21.59 b	1.19-1.59	1.36 b
	C	16.74-18.34	17.54 b	13.96-14.42	14.19 a	1.20-1.27	1.24 a
抽薹期 Bolting stage	B	23.20-46.43	34.29 ab	19.70-34.40	25.82 a	1.15-1.61	1.33 b
抽薹期 Bolting stage	C	17.62-19.58	18.60 a	13.82-15.50	14.66 a	1.26-1.28	1.27 a
注：不同类型淀粉粒进行方差分析，不同小写字母表示差异显著（P<0.05），相同字母者表示差异不显著（P>0.05）。 Note: Variance analysis of different types of starch grains, different lowercase letters indicate significant differences(P <0.05）, the same letters indicate that the difference is not significant(P >0.05）.

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