发酵稻壳通过增强氮代谢途径影响烟草幼苗的生长发育

doi:10.3969/j.issn.1004-1524.20230123

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (2): 237-246.DOI: 10.3969/j.issn.1004-1524.20230123

发酵稻壳通过增强氮代谢途径影响烟草幼苗的生长发育

张斌¹(), 袁志辉¹, 彭陆军², 周向平³, 周德英³, 王锡春³^,^*()

1.湖南科技学院湖南省银杏工程技术研究中心,湖南永州 425199
2.永州市思源农业科技有限公司,湖南永州 425000
3.湖南省烟草公司永州市公司,湖南永州 425000

收稿日期:2023-02-09 出版日期:2024-02-25 发布日期:2024-03-05
作者简介:张斌(1981—),男,湖南永州人,博士,副教授,研究方向为植物发育生物学。E-mail:zhangbin27104@163.com
通讯作者: *王锡春,E-mail:530946968@qq.com
基金资助:
湖南省自然科学基金(2022JJ30274);永州市烟草专卖局科技项目(YYJ201915-24)

Fermented rice husk affects the growth and development of tobacco seedlings by enhancing nitrogen metabolism pathway

ZHANG Bin¹(), YUAN Zhihui¹, PENG Lujun², ZHOU Xiangping³, ZHOU Deying³, WANG Xichun³^,^*()

1. Hunan Provincial Engineering Research Center for Ginkgo Biloba, Hunan University of Science and Engineering, Yongzhou 425199, Hunan, China
2. Yongzhou Siyuan Agricultural Technology Co., Ltd., Yongzhou 425000, Hunan, China
3. Yongzhou Branch of Hunan Provincial Tobacco Company, Yongzhou 425000, Hunan, China

Received:2023-02-09 Online:2024-02-25 Published:2024-03-05

摘要/Abstract

摘要：

为探究发酵稻壳替代炭化稻壳在烟草(Nicotiana tabacum L.)漂浮育苗中的可行性,以商用基质为对照,用不同比例发酵稻壳(fermented rice husk, FRH)替换湘烟7号栽培基质中的炭化稻壳(carbonized rice husk, CRH),并在温室中漂浮育苗60 d,测定幼苗形态指标;选取长势最好处理组FRH1,测定根系和生理指标,进行转录组测序,并对相关代谢通路和关键基因进行鉴定和分析。结果表明,6个处理间整齐度差异不大,但FRH处理生长势优于CRH处理;综合比较,FRH1处理在株高、根冠比和壮苗指数方面优于其他处理。FRH1处理的一级侧根数量、长度和表面积分别为292条、39.7 mm、4.9 cm²,根系脯氨酸含量、CAT和SOD活性分别为7.0 μg·g^-1、232 U·g^-1·min^-1和45.7 U·g^-1·min^-1,均显著高于CRH处理;FRH1和CRH处理共有294个显著差异表达基因参与氮代谢、戊糖和葡萄糖醛酸相互转化等20个KEGG代谢通路,并得到了显著富集;共挖掘出12个参与氮代谢的关键基因,并通过qRT-PCR进行了验证,同时发现硝酸盐/亚硝酸盐转运体基因newGene_31743在2个处理(FRH1和CRH处理)中相对表达量都较高。与CRH处理相比,FRH1处理中烟苗根系硝酸还原酶、亚硝酸还原酶活性分别显著提高8%和7%,氮含量上升1.7%(无显著差异)。发酵稻壳替代炭化稻壳进行漂浮育苗可以增强烟草幼苗氮代谢途径,从而改善烟草根系酶活性,提高根系活力,改良根系形态,进而促进烟苗生长。

关键词: 发酵稻壳, 炭化稻壳, 漂浮育苗, 烟草幼苗

Abstract:

In order to explore the feasibility of replacing carbonized rice husk (CRH) with fermented rice husk (FRH) in floating seedling cultivation of tobacco (Nicotiana tabacum L.). In this study, FRH was used to replace the CRH in different proportions in the culture medium of tobacco with commercial substrate as the control, and the tobacco seedlings were floated in the greenhouse for 60 days, and the morphological indexes of the seedlings were measured. The root and physiological indicators of the best growing treatment group FRH1 were measured, the transcriptome was sequenced, and the related metabolic pathways were analyzed, at the same time, the related key genes of the tobacco seedlings were identified and verified. The results showed that there was no significant difference in uniformity among the six treatments, but the growth potential of FRH treatment was better than that of CRH treatment; By comprehensive comparison, FRH1 treatment was superior to other treatments in plant height, root-shoot ratio and seedling strength index. The number, length and surface area of the primary lateral roots of tobacco seedlings treated with FRH1 were 292, 39.7 mm and 4.9 cm², and the proline content, CAT and SOD activity in root system were 7.0 μg·g^-1, 232 U·g^-1·min^-1 and 45.7 U·g^-1·min^-1, respectively, which were significantly higher than the CRH treatment. Transcriptome data showed that 294 differentially expressed genes in two treatment were significantly enriched in 20 KEGG metabolic pathways, including nitrogen metabolism, pentose and glucuronic acid conversion. 12 key genes involved in nitrogen metabolism pathway were excavated and verified by qRT-PCR analysis, and the relative expression of nitrate/nitrite transporter gene newGene_31743 was higher in both treatments (FRH1 and CRH). The activities of nitrate reductase and nitrite reductase in the roots of tobacco seedlings treated with FRH1 increased by 8% and 7% respectively compared to the CRH treatment, showing significant differences; while the nitrogen content increased by 1.7%, but there was no significant difference. The floating seedling cultivation with FRH instead of CRH can enhance the nitrogen metabolism pathway of tobacco seedlings, thereby improving the enzyme activity of tobacco roots, increasing root activity, improving root morphology, and promoting the growth of tobacco seedlings.

Key words: fermented rice husk, carbonized rice husk, floating nursery, tobacco seedling

中图分类号:

S572

张斌, 袁志辉, 彭陆军, 周向平, 周德英, 王锡春. 发酵稻壳通过增强氮代谢途径影响烟草幼苗的生长发育[J]. 浙江农业学报, 2024, 36(2): 237-246.

ZHANG Bin, YUAN Zhihui, PENG Lujun, ZHOU Xiangping, ZHOU Deying, WANG Xichun. Fermented rice husk affects the growth and development of tobacco seedlings by enhancing nitrogen metabolism pathway[J]. Acta Agriculturae Zhejiangensis, 2024, 36(2): 237-246.

图/表 9

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处理 Treatment	体积分数Volume fraction
处理 Treatment	发酵稻壳 Fermented rice husk	炭化稻壳 Carbonized rice husk	草炭 Turf	珍珠岩 Perlite	蛭石 Vermiculite	牛粪 Cow dung
FRH1	40	0	27.5	20	7.5	5
FRH2	30	10	27.5	20	7.5	5
FRH3	20	20	27.5	20	7.5	5
FRH4	10	30	27.5	20	7.5	5
FRH5	5	35	27.5	20	7.5	5
CRH	0	40	27.5	20	7.5	5

处理 Treatment	体积分数Volume fraction
处理 Treatment	发酵稻壳 Fermented rice husk	炭化稻壳 Carbonized rice husk	草炭 Turf	珍珠岩 Perlite	蛭石 Vermiculite	牛粪 Cow dung
FRH1	40	0	27.5	20	7.5	5
FRH2	30	10	27.5	20	7.5	5
FRH3	20	20	27.5	20	7.5	5
FRH4	10	30	27.5	20	7.5	5
FRH5	5	35	27.5	20	7.5	5
CRH	0	40	27.5	20	7.5	5

基因ID Gene ID	正向引物序列(5'→3') Forward primer sequence (5'→3')	反向引物序列(5'→3') Reverse primer sequence(5'→3')
newGene_21354	ATTGCCGTGGCCTTCCCTT	AGCTGCCAATGGGACCTTC
newGene_24233	ATCACTTTGTGTTGTGAGACTGA	AGAATCTTCCTTGTTGTAGTGAAAG
newGene_2614	CCGAAGCTGAGAGGCAGAAA	AGCAAATAGGAAAAAGTCAGACACG
newGene_2616	GCAGAGTACACCGAAGCTGA	AGCAAATAGAAGAAAGTCAGACACG
newGene_26504	ACTCCGCCTAATGTAACGCC	GCACACGTTTACTATGAGGATGG
newGene_31743	AAACAGCTTGAAGTTCGCCG	TGAGACTCAGCTCATTGAATCTT
gene_21870	TTGCGGGCAAGCCATAATTG	ATCCTCACCGGCTTTGTCAC
gene_31399	GAGAGATCCGGCTTACTGTGG	AGTCTCGATTATGGCTTGTCCA
gene_41133	GCGGGCAAGCCATAATTGAG	CATCCTCACAGGCTGCGTTA
gene_53574	ACGATCGATGAGGTGGGGTA	TCGAGAAATGACCCTAACGGC
gene_56873	GTGCAAGACTCACAACTGCG	AGGCTTCCCATCCCCTTAGT
gene_73099	ACCCCGGTGGTTCTGACA	AGGTGCTAGAAAGCTGCTGA
gene_1665	TGCCCCTGTAATCTCAGCAG	TAGCCACAGCAGCATTGACA

基因ID Gene ID	正向引物序列(5'→3') Forward primer sequence (5'→3')	反向引物序列(5'→3') Reverse primer sequence(5'→3')
newGene_21354	ATTGCCGTGGCCTTCCCTT	AGCTGCCAATGGGACCTTC
newGene_24233	ATCACTTTGTGTTGTGAGACTGA	AGAATCTTCCTTGTTGTAGTGAAAG
newGene_2614	CCGAAGCTGAGAGGCAGAAA	AGCAAATAGGAAAAAGTCAGACACG
newGene_2616	GCAGAGTACACCGAAGCTGA	AGCAAATAGAAGAAAGTCAGACACG
newGene_26504	ACTCCGCCTAATGTAACGCC	GCACACGTTTACTATGAGGATGG
newGene_31743	AAACAGCTTGAAGTTCGCCG	TGAGACTCAGCTCATTGAATCTT
gene_21870	TTGCGGGCAAGCCATAATTG	ATCCTCACCGGCTTTGTCAC
gene_31399	GAGAGATCCGGCTTACTGTGG	AGTCTCGATTATGGCTTGTCCA
gene_41133	GCGGGCAAGCCATAATTGAG	CATCCTCACAGGCTGCGTTA
gene_53574	ACGATCGATGAGGTGGGGTA	TCGAGAAATGACCCTAACGGC
gene_56873	GTGCAAGACTCACAACTGCG	AGGCTTCCCATCCCCTTAGT
gene_73099	ACCCCGGTGGTTCTGACA	AGGTGCTAGAAAGCTGCTGA
gene_1665	TGCCCCTGTAATCTCAGCAG	TAGCCACAGCAGCATTGACA

处理 Treatment	一级侧根Primary lateral root				脯氨酸含量 Proline content/ (μg·g^-1)	过氧化氢酶活性 CAT activity/ (U·g^-1·min^-1)	超氧化物歧化酶活性 SOD activity/ (U·g^-1min^-1)	丙二醛含量 MDA content/ (mmol·g^-1)
处理 Treatment	数量 Quantity	长度 Length/ mm	直径 Diameter/ mm	表面积 Surface area/cm²	脯氨酸含量 Proline content/ (μg·g^-1)	过氧化氢酶活性 CAT activity/ (U·g^-1·min^-1)	超氧化物歧化酶活性 SOD activity/ (U·g^-1min^-1)	丙二醛含量 MDA content/ (mmol·g^-1)
FRH1	292±32 a	39.7±2.7a	0.066±0.003 a	4.9±0.5 a	7.0±1.6 a	232±8 a	45.7±4.5 a	1.5±0.4 a
CRH	194±35 b	28.3±3.7b	0.063±0.009 a	3.2±0.4 b	10.6±1.6 b	204±5 b	27.1±9.5 b	1.8±0.3 a

发酵稻壳通过增强氮代谢途径影响烟草幼苗的生长发育

Fermented rice husk affects the growth and development of tobacco seedlings by enhancing nitrogen metabolism pathway

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处理 Treatments	过滤序列 Filter sequence	Q30/%	GC含量 GC content/%	比对数 Comparison number	比对率 Comparison rate/%	单一比对率 Single comparison rate/%
FRH1-1	48 113 920	94.67	43.22	44 752 999	95.20	93.01
FRH1-2	45 998 192	94.44	43.16	42 797 732	95.19	93.04
FRH1-3	51 248 126	94.68	43.12	47 770 763	95.42	93.21
CRH 1	45 101 078	94.29	43.28	41 752 391	94.80	92.58
CRH 2	43 370 834	94.78	43.31	40 360 157	95.29	93.06
CRH 3	54 891 242	94.48	43.35	50 322 002	93.89	91.68

基因ID Gene ID	功能注释 Annotation	表达水平 Expression level	RNA-Seq(FPKM) log₂(fold-change)	相对表达量(FRH1/CRH) Relative expression
newGene_21354	硝酸盐转运蛋白Nitrate transporter	下调Down-regulated	-1.89	0.29
newGene_24233	硝酸盐转运蛋白Nitrate transporter	下调Down-regulated	-1.06	0.49
newGene_2614	硝酸盐转运蛋白Nitrate transporter	上调Up-regulated	0.64	1.60
newGene_2616	硝酸盐转运蛋白Nitrate transporter	上调Up-regulated	0.60	1.56
newGene_26504	硝酸盐转运蛋白Nitrate transporter	下调Down-regulated	-1.26	0.43
newGene_31743	硝酸盐转运蛋白Nitrate transporter	上调Up-regulated	0.72	1.69
gene_21870	亚硝酸盐还原酶Nitrite reductase	上调Up-regulated	1.01	2.07
gene_31399	亚硝酸盐还原酶Nitrite reductase	上调Up-regulated	0.87	1.87
gene_41133	亚硝酸盐还原酶Nitrite reductase	上调Up-regulated	0.81	1.79
gene_53574	硝酸还原酶Nitrate reductase	上调Up-regulated	1.16	2.28
gene_73099	硝酸还原酶Nitrate reductase	上调Up-regulated	0.77	1.74
gene_56873	谷氨酸脱氢酶Glutamate dehydrogenase	上调Up-regulated	0.67	1.77