花魔芋球茎发育期氮分配变化和相关基因表达分析

doi:10.16420/j.issn.0513-353x.2023-0629

摘要/Abstract

摘要：

利用¹⁵N同位素示踪和转录组测序，系统地研究了花魔芋（Amorphophallus konjac K. Koch）球茎发育过程中各器官氮素分配特征，以及氮代谢相关基因表达情况，以理解氮素利用情况并指导合理用肥。结果表明，在球茎发育过程中，叶片的含氮量最高；随球茎膨大，叶片的含氮量、Ndff值、¹⁵N分配率和利用率呈下降趋势，而球茎对氮素的需求增加，¹⁵N利用率高峰出现在球茎膨大中后期（散叶后45 ~ 75 d）。整个植株¹⁵N利用率呈现先增加后降低的趋势，在散叶后60 d时整个植株中的N含量达到峰值，¹⁵N利用率最高，为22.68%，之后在各个器官中N素含量与¹⁵N利用率均逐步降低，说明在60 d时是氮素的最大利用效率期，从此开始植物吸收的氮素由主要分配给叶片转向分配给球茎，此时为追施氮肥的适宜期。叶片中氮代谢相关基因表达高峰出现在球茎膨大中后期（散叶后105 d），此时叶片的氮代谢反应加强，有利于氮同化产物合成和输出。叶片中硝酸还原酶基因AkNIA1和谷氨酰胺合成酶基因AkGSR2-1表达水平与叶片和根中N含量显著正相关，表明其在促进氮吸收和氮积累中的重要作用，可作为氮利用率遗传改良的候选基因。

关键词: 花魔芋, 氮素分配, 氮代谢, 基因表达, 球茎, ¹⁵N同位素示踪

Abstract:

Through ¹⁵N isotopic tracing and RNA-seq analysis，the N distribution in various organs and the expression of genes related to N during the development of konjac corms are systematically studied would help to understand nitrogen utilization and guide the fertilizer use. Results show that during the development of Amorphophallus konjac corms the N content per unit weight of leaves was the highest as the compared with other organs. During the expansion of corms，N content，Ndff value，¹⁵N distribution rate and ¹⁵N utilization rate in leaves showed a downward trend，whereas the N demand in corms increased，the peak of ¹⁵N utilization rate appearing right in the middle stage of corm expansion（45-75 d，day after expansion of leaves）. A. konjac reached the peak of ¹⁵N content of in a whole plant at 60 d，while the ¹⁵N utilization rate also reached the highest rate of 22.68%. Then，both the content and utilization rate of ¹⁵N were deceasing，indicating that this point was the most efficiently nitrogen use period. After 60 d，the absorbed nitrogen was distributed from leaves to corms，which also invalided that 60 d was the suitable period for N topdressing. Meanwhile，the peak of expression in N metabolism-related genes in leaves appeared between the middle and late stage of corm expansion（105 d），a range within which leaf N metabolism was strengthened，promoting the synthesis and output of N assimilation products. Among them，the expression levels in leaves of nitrate reductase gene AkNIA1 and glutamine synthetase gene AkGSR2-1 were significantly and positively correlated with the N content in leaves and roots，indicating their key role in nitrogen absorption and accumulation，and they can be used as candidate genes for genetic improvement of N utilization.

Key words: Amorphophallus konjac, nitrogen distribution, nitrogen metabolism, gene expression, corm, ¹⁵N isotopic tracing

任思源, 陈森, 龙治坚, 王博雅, 唐登国, 王正前, 杨斌, 胡尚连, 曹颖. 花魔芋球茎发育期氮分配变化和相关基因表达分析[J]. 园艺学报, 2024, 51(9): 2019-2030.

REN Siyuan, CHEN Sen, LONG Zhijian, WANG Boya, TANG Dengguo, WANG Zhengqian, YANG Bin, HU Shanglian, CAO Ying. Nitrogen Allocation Characteristics and Expression of Related Genes During Corm-Forming Stage of Amorphophallus konjac[J]. Acta Horticulturae Sinica, 2024, 51(9): 2019-2030.

图/表 10

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引物名称 Primer name	引物序列（5′-3′） Primer sequence	引物名称 Primer name	引物序列（5′-3′） Primer sequence
AkNIA1-F	GTGTGGTACGTGGTGAATGAG	AkGSR2-1-F	TGGAACTATGATGGATCGAGCA
AkNIA1-R	GTTCTTAACGTCGTAGCCCATC	AkGSR2-1-R	TGTGTACTCCTGTTCAATCCCA
AkNIR1-F	GAAGGTGAAGCTGGAGAAGGAG	AkASN1-F	AGAGAATTTCGTGGACATGCTG
AkNIR1-R	CTTCAGCCTCATCATGAACCG	AkASN1-R	AAATGCTCGCAGTCATCATTGA
AkGLU1-F	GTTGTCCTTGGAAAAGTGGGAA	Tubulin-F	GCCGTGAATCTCATCCCCTT
AkGLU1-R	ATCCTGCCCATCACAAATTCTG	Tubulin-R	TTGTTCTTGGCATCCCACAT

引物名称 Primer name	引物序列（5′-3′） Primer sequence	引物名称 Primer name	引物序列（5′-3′） Primer sequence
AkNIA1-F	GTGTGGTACGTGGTGAATGAG	AkGSR2-1-F	TGGAACTATGATGGATCGAGCA
AkNIA1-R	GTTCTTAACGTCGTAGCCCATC	AkGSR2-1-R	TGTGTACTCCTGTTCAATCCCA
AkNIR1-F	GAAGGTGAAGCTGGAGAAGGAG	AkASN1-F	AGAGAATTTCGTGGACATGCTG
AkNIR1-R	CTTCAGCCTCATCATGAACCG	AkASN1-R	AAATGCTCGCAGTCATCATTGA
AkGLU1-F	GTTGTCCTTGGAAAAGTGGGAA	Tubulin-F	GCCGTGAATCTCATCCCCTT
AkGLU1-R	ATCCTGCCCATCACAAATTCTG	Tubulin-R	TTGTTCTTGGCATCCCACAT

散叶后天数/d Days after expansion of leaves	叶片 Leaf	叶柄 Petiole	根 Root	球茎 Corm
30	35.33 ± 0.83 a	15.58 ± 1.09 c	29.00 ± 3.95 a	12.21 ± 0.96 c
45	34.90 ± 0.58 a	19.02 ± 1.59 b	28.84 ± 0.79 a	21.06 ± 5.56 a
60	35.68 ± 2.11 a	21.79 ± 1.04 a	29.36 ± 4.71 a	17.25 ± 0.14 abc
75	29.19 ± 0.71 b	14.55 ± 1.15 c	26.47 ± 2.44 a	15.09 ± 2.19 bc
105	23.84 ± 1.30 c	15.69 ± 0.94 c	23.81 ± 2.78 a	17.95 ± 1.51 ab

散叶后天数/d Days after expansion of leaves	叶片 Leaf	叶柄 Petiole	根 Root	球茎 Corm
30	35.33 ± 0.83 a	15.58 ± 1.09 c	29.00 ± 3.95 a	12.21 ± 0.96 c
45	34.90 ± 0.58 a	19.02 ± 1.59 b	28.84 ± 0.79 a	21.06 ± 5.56 a
60	35.68 ± 2.11 a	21.79 ± 1.04 a	29.36 ± 4.71 a	17.25 ± 0.14 abc
75	29.19 ± 0.71 b	14.55 ± 1.15 c	26.47 ± 2.44 a	15.09 ± 2.19 bc
105	23.84 ± 1.30 c	15.69 ± 0.94 c	23.81 ± 2.78 a	17.95 ± 1.51 ab

散叶后天数/d Days after expansion of leaves	叶片 Leaf	叶柄 Petiole	根 Root	球茎 Corm	全株 Whole plant
30	1.17 ± 0.04 a	1.12 ± 0.10 ab	1.12 ± 0.06 ab	0.89 ± 0.11 b	4.31 ± 0.19 a
45	0.87 ± 0.22 b	1.34 ± 0.17 a	1.06 ± 0.16 b	1.22 ± 0.04 a	4.49 ± 0.58 a
60	0.85 ± 0.12 b	1.36 ± 0.16 a	1.27 ± 0.09 a	1.34 ± 0.08 a	4.81 ± 0.44 a
75	0.59 ± 0.11 c	0.87 ± 0.17 b	0.55 ± 0.02 d	0.86 ± 0.12 b	2.88 ± 0.38 b
105	0.56 ± 0.04 c	0.90 ± 0.05 b	0.78 ± 0.06 c	0.60 ± 0.02 c	2.84 ± 0.07 b