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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (5): 807-813.doi: 10.3724/SP.J.1006.2021.03039

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Function analysis of nitrogen-responsive transcription factor ZmNLP5 affecting root growth in maize

GE Min(), WANG Yuan-Cong, NING Li-Hua, HU Meng-Mei, SHI Xi, ZHAO Han*()   

  1. Institute of Crop Germplasm and Biotechnology / Provincial Key Laboratory of Agrobiology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu, China
  • Received:2020-06-20 Accepted:2020-11-13 Online:2021-05-12 Published:2020-12-15
  • Contact: ZHAO Han E-mail:gemin8614@163.com;zhaohan@jaas.ac.cn
  • Supported by:
    National Natural Science Foundation of China(32001564);China Key Research and Development Program of Jiangsu Province(BE207365);Jiangsu Agriculture Science and Technology Innovation(CX(18)1001)

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Abstract:

Improving nitrogen (N) use efficiency of crops is crucial for minimizing N loss and reducing environmental pollution, which is a requirement for the sustainable agriculture. Our previous study found that the transcription factor ZmNLP5 directly regulated the expression of ZmNIR1.1 and promoted nitrogen uptake and assimilation in maize, however, its underlying regulation mechanism is unclear. Here, we performed further phenotype analysis of zmnlp5 and wild type (W22) plants in hydroponic culture on sufficient nitrogen (SN) solution and deficient nitrogen (DN) solution. Compared with WT plants, the root length of zmnlp5 mutant plants was significantly decreased under DN condition. Compared with upper and middle regions of roots, ZmNLP5 was predominantly expressed in root tip regions. Then, we examined the relationship between root length and nitrite content in root tips under a series of nitrite concentrations. The results showed that there was no significant differences in root length between WT and zmnlp5 until the nitrite concentration reached 2 mmol L-1 and higher; however, when the concentration of nitrite was higher than 2 mmol L-1, the root length of zmnlp5 was significantly shorter than WT, and the accumulation of nitrite in the root tips of zmnlp5 was significantly higher than WT. Interestingly, zmnlp5 plants also accumulated significantly more nitrite in the root tips than WT under DN condition. The study showed that the transcription factor ZmNLP5 played an important role in the root growth of maize in response to deficient nitrogen condition, and provided the candidate genes for breeding of maize nitrogen use efficiency in the future.

Key words: maize, nitrogen, transcription factor, nitrogen response, root length

Fig. 1

Root length analysis of WT and zmnlp5 mutants on SN and DN solution in hydroponic culture A: phenotype analysis (SN: sufficient nitrogen; DN: deficient nitrogen; Bar = 20 mm); B: root length analysis. **: P ≤ 0.01 by Student’s t-test (n = 3)."

Fig. 2

Relative expression of ZmNLP5 in different root regions A: mRNA relative expression levels were measured by qPCR and normalized to ZmUPF1. Different letters marked above the bars indicate significant differences at P ≤ 0.05; B: protein expression levels were measured by immunoblot assay using anti-UDPGP as a sample loading control."

Fig. 3

Root length analysis of WT and zmnlp5 mutants under nitrite treatment in hydroponic culture A: phenotypes of WT and zmnlp5 plants grown in hydroponic culture for seven days on DN solution with 0, 0.5, 1, 2, and 5 mmol L-1 KNO2 for seven days. Bar = 20 mm; B: root length of WT and zmnlp5 plants grown in hydroponic culture for seven days on DN solution with 0, 0.5, 1, 2, and 5 mmol L-1 KNO2 for seven days. * P ≤ 0.05 by Student’s t-test (n = 3)."

Fig. 4

Nitrite content in root tips of WT and zmnlp5 mutants under nitrite treatments ** P ≤ 0.01, t测验, n = 3。** P ≤ 0.01 by Student’s t-test (n = 3)."

Fig. 5

Nitrite content in different regions of the roots of WT and zmnlp5 mutants on SN solution and DN solution in hydroponic culture SN: sufficient nitrogen; DN: deficient nitrogen. * P ≤ 0.05 by Student’s t-test (n = 3)."

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