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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (5): 1372-1385.doi: 10.3724/SP.J.1006.2023.24133

• TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

Effects of exogenous tryptophan on root elongation of sorghum seedlings under low nitrogen stress

LI Bang(), LIU Chun-Juan(), GUO Jun-Jie, WU Yu-Xin, DENG Zhi-Cheng, ZHANG Min, CUI Tong, LIU Chang, ZHOU Yu-Fei()   

  1. College of Agronomy, Shenyang Agricultural University, Shenyang 110866, Liaoning, China
  • Received:2022-06-01 Accepted:2022-09-05 Online:2023-05-12 Published:2022-09-22
  • Contact: *E-mail: zhouyufei@syau.edu.cn
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    China Agriculture Research System of MOF and MARA(CARS-06-14.5-A17);General Research Project of Education Department of Liaoning Province of China(LSNFW202006)

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Abstract:

The physiological mechanism of root elongation in sorghum under low nitrogen stress remains unclear. Here, two sorghum inbred lines, 398B (low nitrogen tolerance) and CS-3541 (low nitrogen sensitivity), were used as experimental materials, to clarify the physiological mechanism of root elongation of sorghum seedlings under low nitrogen stress. The results showed that, compared with normal nitrogen stress, low nitrogen stress significantly increased root length and root tip cell length of 398B and CS-3541, and 398B had longer root length. The endogenous tryptophan content in roots of 398B and CS-3541 increased significantly at 1, 5, and 10 days after low nitrogen stress. Tryptophan was involved in root elongation of sorghum under low nitrogen conditions through the auxin synthesis pathway by RNA-seq, and the relative expression level of genes in the auxin synthesis pathway of root in 398B was significantly higher than that in CS-3541. Furthermore, the root lengths of 398B and CS-3541 were significantly increased by 50 mg L-1 exogenous tryptophan treatment under low nitrogen conditions. Exogenous tryptophan activated the activity of H+-ATPase in plasma membrane by increasing the content of auxin, promoted the acidification of plasma membrane, and improved the activity of enzymes related to energy metabolism and ATP content, and thus inducing energy metabolism of root system. Exogenous tryptophan had a better effect on 398B under low nitrogen stress. In conclusion, low nitrogen stress activated the key role of endogenous tryptophan in sorghum root elongation, and auxin depended on tryptophan pathway and synergistic enhancement of energy metabolism were the physiological mechanism promoting root elongation of sorghum seedlings under low nitrogen stress.

Key words: sorghum, low nitrogen, exogenous tryptophan, auxin, energy metabolism, root elongation

Table 1

Sequence primer for qPCR used in this study"

基因 Gene name PCR引物 Primer sequences (5°-3°)
YUCCA10 F: GATGGATGTGAGGAGCAAAG; R: TGAACGGGTCTCTGAAGAT
YUCCA9 F: CATCATCCGTGAGAACCTTG; R: ATGATGGCCATGCAGAAC
TAA F: AGAGCATCCGTCTCTTCTC; R: TGGTGAGGTTGAGGTTGT

Fig. 1

Root phenotype (A) and length (B) of sorghum seedling under low nitrogen stress Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment."

Fig. 2

Root tip cell morphology (A) and length (B) of sorghum seedlings under low nitrogen stress Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment."

Fig. 3

Effect of low nitrogen stress on root tryptophan content of sorghum seedlings I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment."

Fig. 4

Tryptophan metabolic pathway"

Fig. 5

Effects of low nitrogen stress on auxin synthesis-related gene expression levels in sorghum seedlings Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment."

Fig. 6

Effect of exogenous tryptophan on root growth of sorghum seedlings under low nitrogen stress NN: normal nitrogen treatment; LN: low nitrogen treatment."

Fig. 7

Effects of exogenous tryptophan on auxin content in roots of seedlings under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 8

Effects of exogenous tryptophan on plasma membrane H+-ATPase activity in roots of sorghum seedlings under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 9

Effects of exogenous tryptophan on root ectoplasmic pH of sorghum seedlings under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 10

Effects of exogenous tryptophan on ATP content of sorghum seedling roots under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 11

Effects of exogenous tryptophan on glucose-6-phosphate dehydrogenase activity in roots of sorghum seedlings under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 12

Effects of exogenous tryptophan on pyruvate kinase activity in sorghum seedling roots under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 13

Effects of exogenous tryptophan on citrate synthase activity in roots of sorghum seedlings under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 14

Effects of exogenous tryptophan on α-ketoglutarate dehydrogenase activity in roots of sorghum seedling under low nitrogen stress I: 1 day after low nitrogen treatment; II: 5 days after low nitrogen treatment; III: 10 days after low nitrogen treatment. C, S, and B represent hybrid lines, different nitrogen treatments, and exogenous tryptophan treatments, respectively. *, **, and *** are significantly different at the 0.05, 0.01, and 0.001 probability levels, respectively, and ns is not significantly different. Columns with different lowercase letters are significantly different at the 0.05 probability level. NN: normal nitrogen treatment; LN: low nitrogen treatment; NN-T: normal nitrogen was sprayed with tryptophan; LN-T: low nitrogen was sprayed with tryptophan."

Fig. 15

Model of exogenous tryptophan on root elongation of sorghum seedlings under low nitrogen stress"

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