细胞分裂素参与硝酸盐介导苹果不定根发生的生理和分子机制研究

doi:10.1016/j.jia.2024.07.045

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (12): 4046-4057.DOI: 10.1016/j.jia.2024.07.045

所属专题：园艺作物基因功能与分子调控机制Horticulture — Gene function · Molecular mechanism

细胞分裂素参与硝酸盐介导苹果不定根发生的生理和分子机制研究

收稿日期:2023-08-17 接受日期:2024-06-17 出版日期:2024-12-20 发布日期:2024-11-15

Physiological and molecular mechanisms of cytokinin involvement in nitrate-mediated adventitious root formation in apples

Muhammad Mobeen Tahir^1*, Li Fan^1*, Zhimin Liu¹, Humayun Raza², Usman Aziz³, Asad Shehzaib¹, Shaohuan Li¹, Yinnan He¹, Yicen Lu¹, Xiaoying Ren¹, Dong Zhang^1#, Jiangping Mao^1#

¹College of Horticulture, Yangling Sub-Center of National Center for Apple Improvement, Northwest A&F University, Yangling 712100, China
²Department of Plant Breeding and Genetics, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur, Bahawalpur 63100, The Islamic Republic of Pakistan
³National Cotton Breeding Institute, The Islamia University of Bahawalpur, Bahawalpur 63100, The Islamic Republic of Pakistan

Received:2023-08-17 Accepted:2024-06-17 Online:2024-12-20 Published:2024-11-15
About author:#Correspondence Jianping Mao, E-mail: mjp588@nwafu.edu.cn; Dong Zhang, E-mail: afant@nwsuaf.edu.cn *These authors contributed equally to this study.
Supported by:
This work was financially supported by the National Natural Science Foundation of China (32372675, 32372657, 32102359), the National Key Research and Development Project, China (2023YFD2301002), the Young Talent Fund of Association for Science and Technology in Shaanxi, China (20240218), the Science and Technology Major Project of Xinjiang Production and Construction Corps, China (2023AB077), the Chinese Universities Scientific Fund (2452023005), the China Apple Research System (CARS-27), the Cyrus Tang Foundation, and the Fundamental Research Funds for the Central Universities, China.

摘要/Abstract

摘要：

硝酸盐被认为促进苹果不定根（AR）的形成，但细胞分裂素 (CK) 是否参与硝酸盐介导的不定根形成尚不明确。本试验以苹果GL-3组培苗为材料，对其进行不同硝酸盐组合处理。结果显示，T1 (硝酸盐0.95 g/L + 6-BA 0.5 mg/L) 和T3 (6-BA 0.5 mg/L) 处理显著抑制不定根的形成。T2 (硝酸盐0.95 g/L) 和T4 (硝酸盐0.95 g/L +洛伐他汀 0.5 mg/L) 处理均促进不定根的形成。然而，T4产生的不定根较少且短，说明适量的CK对不定根的正常发育是必需的，这也表明，这些不定根是由于硝酸盐的存在而产生的。进一步解剖观察发现，抑制CK生物合成延迟了不定根原基的形成。T2处理显著提高了其内源IAA和ZR的含量，而T4处理后内源ABA、GA和BR含量较高。同时，在T2处理后，MdNRT1.1和MdNRT2.1在第3天和第8天表达量较高，而MdRR2和MdCKX5在第8天和第16天表达量较高。此外，较高的IAA水平诱导MdWOX11的表达，从而上调MdLBD16和MdLBD29的表达。这些基因的联合表达通过上调细胞周期相关基因 (MdCYCD1;1和MdCYCD3;1) 来诱导不定根的发生。综上，本研究发现特定的基因和激素在硝酸盐- CK介导的苹果不定根的发生中发挥重要作用。

Abstract:

Potassium nitrate (KNO₃) promotes adventitious root (AR) formation in apple stem cuttings. However, evidence for the possible involvement of cytokinin (CK) in KNO₃-mediated AR formation in apples is still lacking. In this study, we cultured GL-3 apple microshoots in different treatment combinations. While the T1 (KNO₃ 9.4 mmol L^–1+6-benzyl adenine (6-BA) 2.22 μmol L^–1) and T3 (6-BA 2.22 μmol L^–1) treatments completely inhibited AR formation, the control, T2 (KNO₃ 9.4 mmol L^–1), and T4 (KNO₃ 9.4 mmol L^–1+lovastatin (Lov) 1.24 μmol L^–1) treatments developed ARs. However, T4-treated microshoots developed fewer and shorter ARs, indicating that optimum CK synthesis is needed for normal AR growth. This also suggests that these fewer and shorter ARs developed because of the presence of KNO₃ in the same medium. The anatomy of the stem basal part indicated that the inhibition of CK biosynthesis delayed AR primordia formation. The endogenous levels of indole‐3‐acetic acid (IAA) and zeatin riboside (ZR) were higher in T2-treated microshoots, while the abscisic acid (ABA), gibberellic acid 3 (GA₃), and brassinosteroid (BR) levels were higher in T4-treated microshoots. The expression levels of MdNRT1.1 and MdNRT2.1 were higher in T2-treated microshoots at 3 and 8 days, while MdRR2 and MdCKX5 were higher at 8 and 16 days, respectively. Furthermore, higher IAA levels increased MdWOX11 expression, which in turn increased MdLBD16 and MdLBD29 expression in response to T2. The combined expression of these genes stimulated adventitious rooting by upregulating cell cycle-related genes (MdCYCD1;1 and MdCYCD3;1) in response to T2 treatment. This study shows that specific genes and hormonal pathways contribute to KNO₃-CK-mediated adventitious rooting in apples.

Key words: adventitious root (AR) , cytokinin (CK) , potassium nitrate (KNO₃) , crosstalk , hormones , gene expression

Muhammad Mobeen Tahir, Li Fan, Zhimin Liu, Humayun Raza, Usman Aziz, Asad Shehzaib, Shaohuan Li, Yinnan He, Yicen Lu, Xiaoying Ren, Dong Zhang, Jiangping Mao. 细胞分裂素参与硝酸盐介导苹果不定根发生的生理和分子机制研究[J]. Journal of Integrative Agriculture, 2024, 23(12): 4046-4057.

Muhammad Mobeen Tahir, Li Fan, Zhimin Liu, Humayun Raza, Usman Aziz, Asad Shehzaib, Shaohuan Li, Yinnan He, Yicen Lu, Xiaoying Ren, Dong Zhang, Jiangping Mao. Physiological and molecular mechanisms of cytokinin involvement in nitrate-mediated adventitious root formation in apples[J]. Journal of Integrative Agriculture, 2024, 23(12): 4046-4057.

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细胞分裂素参与硝酸盐介导苹果不定根发生的生理和分子机制研究

Physiological and molecular mechanisms of cytokinin involvement in nitrate-mediated adventitious root formation in apples

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