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

doi:10.3969/j.issn.1004-1524.20230123

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (2): 237-246.DOI: 10.3969/j.issn.1004-1524.20230123

• Crop Science • Previous Articles Next Articles

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

Abstract

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

CLC Number:

S572

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.

Figures/Tables 9

References 34

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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

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