 |
 |
 |
 |
 |
 |
Sains Malaysiana 54(11)(2025): 2661-2673
http://doi.org/10.17576/jsm-2025-5411-07
Interstock-Induced Anatomical and Metabolomic Shifts Enhance
Growth and Stress Adaptation in Citrus Cultivars
(Perubahan Anatomi dan Metabolomik yang Dipicu oleh Interstok Meningkatkan Tumbesaran dan Penyesuaian terhadap Stres dalam Kultivar Sitrus)
AGUS SUGIYATNO1, NIRMALA FRIYANTI DEVY1,*, HARDIYANTO1, ANANG TRIWIRATNO1,
SRI WIDYANINGSIH1, EMI BUDIYATI1, FARIDA YULIANTI1, HASIM ASHARI1, MUTIA ERTI
DWIASTUTI1, DORKAS PARHUSIP1 & IMRO’AH IKARINI2
1Research Center for Horticulture, National Research and Innovation
Agency of Indonesia, Cibinong Science Center, Jalan Raya Bogor, Cibinong,
Indonesia
2Research Center for Agroindustry, National Research and Innovation
Agency of Indonesia, Kawasan Puspitek, Tangerang Selatan, Banten, Indonesia
Received:
29 October 2025/Accepted: 20 May 2025
Citrus growers are challenged to be able to produce high yields
while being resilient to environmental stress, while maintaining high yield and
cultivar potential. However, the use of pummelo as an interstock-despite its growing relevance for cultivar
replacement-remains poorly studied in terms of its anatomical and metabolic
effects on scion varieties. This study evaluates the impact of Pummelo
interstock on the growth, leaf anatomy, and metabolomic profile of three citrus
cultivars: Sweet Orange (Manis Pacitan), Tangerine (Siam Pontianak), and
Mandarin (Keprok Batu 55). Morphological assessment showed that interstock application increased leaf thickness by 3.9% and
palisade layer thickness by 15.2%, with additional adaptations in epidermal and
glandular structures suggesting enhanced stress tolerance. Stomatal density
rose by 22% in certain combinations. GC-MS metabolomic profiling identified 91
metabolites, with interstock treatments increasing the accumulation of
stress-related compounds such as flavonoids and terpenes. Stress-metabolite
upregulation reached 59.6% in Keprok Batu 55, 36.2% in Manis Pacitan, and 47.8%
in Siam Pontianak. Key pathway modifications involved the stimulation of fatty
acid metabolism and the development of glycosylphosphatidylinositol (GPI)
anchors in interstocked plants, which led to better morphological and
physiological features. These findings highlight the role of interstock technology
in promoting vegetative vigor, metabolic reprogramming, and stress adaptation
in citrus, with practical implications for improving productivity and
sustainability in citrus cultivation. Further research should explore long-term
performance and underlying genetic mechanisms.
Keywords: Citrus; interstock; morpho-anatomical
traits; pummelo; secondary metabolite
Abstrak
Penanam sitrus dicabar untuk menghasilkan hasil yang tinggi dan berdaya tahan terhadap tekanan alam sekitar, sambil mengekalkan potensi
hasil dan kultivar yang tinggi. Walau bagaimanapun, penggunaan pummelo sebagai interstok-walaupun semakin relevan untuk penggantian kultivar-masih kurang dikaji dari segi kesan anatomi dan metaboliknya pada
varieti scion. Kajian ini menilai kesan interstok pumello terhadap pertumbuhan, anatomi daun dan profil metabolomik tiga kultivar sitrus: Jeruk
Manis (Manis Pacitan), Jeruk Keprok (Siam Pontianak) dan Jeruk Mandarin (Keprok
Batu 55). Penilaian morfologi menunjukkan bahawa penggunaan interstok
meningkatkan ketebalan daun sebanyak 3.9% dan lapisan palisade sebanyak 15.2%,
disertai adaptasi pada epidermis dan kelenjar yang mencadangkan peningkatan
toleransi tekanan. Ketumpatan stomata meningkat sebanyak 22% dalam beberapa gabungan. Pemprofilan GC-MS mengenal pasti 91 metabolit dengan interstok meningkatkan pengumpulan sebatian berkaitan tekanan seperti
flavonoid dan terpena. Peningkatan metabolit tekanan dicatatkan sebanyak 59.6%
di Keprok Batu 55, 36.2% di Manis Pacitan dan 47.8%
di Siam Pontianak. Perubahan laluan utama termasuk pengaktifan metabolisme asid lemak dan biosintesis penambat glikosilfosfatidilinositol (GPI) dalam tumbuhan saling berstok, sepadan dengan anatomi dan fisiologi yang lebih baik. Hasil ini
menyerlahkan potensi teknologi interstok dalam meningkatkan kekuatan vegetatif, pengaturcaraan semula metabolik dan penyesuaian tekanan dalam tanaman sitrus.
Penyelidikan lanjut perlu dijalankan untuk menilai prestasi jangka panjang dan
mekanisme genetik yang terlibat.
Kata kunci: Citrus; interstok; ciri morfo-anatomi; metabolit sekunder; pummelo
REFERENCES
Abadie, C., Lalande, J. & Tcherkez, G. 2022. Exact mass GC-MS analysis: Protocol,
database, advantages, and application to plant metabolic profiling. Plant
Cell and Environment 45(10): 3171-3183.
Alves, N.S.F.,
Setzer, W.N. & da Silva, J.K.R. 2019. The chemistry and biological
activities of Peperomia pellucida (Piperaceae):
A critical review. Journal of Ethnopharmacology 232: 90-102.
Anandakumar, P., Kamaraj, S. & Vanitha,
M.K. 2021. D-limonene: A multifunctional compound with potent therapeutic
effects. Journal of Food Biochemistry 45(1): e13566.
Balfagón, D., Terán, F., de Oliveira, T.D.R., Santa-Catarina, C. &
Gómez-Cadenas, A. 2022. Citrus rootstocks modify the scion antioxidant system
under drought and heat stress combination. Plant Cell Reports 41(3): 593-602.
Bennici, S., Las Casas, G., Distefano, G.,
Gentile, A., Lana, G., Di Guardo, M., Nicolosi, E., La Malfa, S. & Continella, A. 2021. Rootstock affects floral induction in
citrus engaging the expression of the FLOWERING LOCUS T (CiFT). Agriculture 11: 140.
Cahyanti, L.D., Sopandie, D., Santosa, E. & Purnamawati,
H. 2024. Diversity of 17 genotypes of taro based on anatomy and nutritional value
of tuber. HAYATI Journal of Biosciences 31(3): 465-473.
Carrera, F.P., Noceda, C., Maridueña-Zavala,
M.G. & Cevallos-Cevallos, J.M. 2021.
Metabolomics, a powerful tool for understanding plant abiotic stress. Agronomy 11(5): 824.
Cookson, S.J.,
Clemente Moreno, M.J., Hevin, C., Nyamba Mendome, L.Z., Delrot, S., Trossat-Magnin, C. & Ollat,
N. 2013. Graft union formation in grapevine induces transcriptional changes
related to cell wall modification, wounding, hormone signalling,
and secondary metabolism. Journal of Experimental Botany 64(10): 2997-3008.
Dah-Nouvlessounon,
D., Chokki, M., Noumavo,
A.D.P., Cârâc, G., Furdui, B., Sina, H., Zongo, C.,
Savadogo, A., Baba-Moussa, L., Dinica, R.M. &
Baba-Moussa, F. 2023. Ethnopharmacological value and biological activities via
antioxidant and anti-protein denaturation activity of Morinda lucida Benth and Momordica charantia L. leaves extracts from Benin. Plants 12(6): 1228.
David, O.A., Labulo,
A.H., Adejayan, M.T., Adeleke, E.A., Adeniyi, I.M.
& Terna, A.D. 2023. Anatomical adaptation of water-stressed Eugenia uniflora using green synthesized silver nanoparticles
and melatonin. Microscopy Research and Technique 86(6): 648-658.
de Carvalho, C.C.C.R. & Caramujo, M.J. 2018. The various roles of fatty acids. Molecules 23(10):
2583. MDPI.
de Lima Costa, M.V., de Lima Félis, P.A., Masselani, K.J.S., Linard, T.L.M., Bucci, L.A. & de Lima Vieira, W. 2021.
Organization of leaf vascular system and gas exchange in seedlings of Guazuma ulmifolia Lam. in different light conditions. Scientific Electronic Archives 14(7): 68-73.
de Lima Vieira, W., Boeger, M.R.T., Cosmo, N.L.
& Coan, A.I. 2014. Leaf morphological plasticity of tree species from two
developmental stages in Araucaria forest. Brazilian Archives of
Biology and Technology 57(4): 476-485.
Deng, H., Achor, D., Exteberria,
E., Yu, Q., Du, D., Stanton, D., Liang, G. & Gmitter, F.G. 2019. Phloem
regeneration is a mechanism for Huanglongbing-tolerance of “Bearss” lemon and
“LB8-9” Sugar Belle® mandarin. Frontiers in Plant Science 10: 277.
Devy, N.F., Hardiyanto, Sugiyatno, A., Dwiastuti, M.E. & Yulianti,
F. 2023. Leaf anatomy, photosynthetic characteristics, fruit quality, and
genetic changes in “Borneo Prima” mandarin (Citrus Reticulata Blanco)
grafted onto different interstocks in dry highland
conditions. Applied Ecology and Environmental Research 21(2): 1805-1822.
Dutt, M., Mahmoud, L.M. & Grosser, J.W.
2023. Field performance of ‘Valencia’ sweet orange trees grafted onto pummelo interstocks and swingle citrumelo rootstocks under Huanglongbing (HLB)
endemic conditions. Horticulturae 9(6): 719.
Dwiastuti,
M.E., Sugiyatno, A., Devy, N.F. & Hardiyanto. 2023. The effect of interstock on the development of Huanglongbing disease and vegetative growth of three
commercial citrus varieties in Indonesia. IOP Conference Series: Earth and
Environmental Science 1172: 012034.
Fayek, M.A., Ali, A.E.M. & Rashedy, A.A. 2022. Physiological and chemical performance
of the Flame seedless grapevine cultivar in the presence of Paulsen 1103 as the interstock. Ciencia e Agrotecnologia 46: e021621.
Fayek, M.A., Rashedy, A.A. & Ali, A.E.M. 2022. Alleviating the
adverse effects of deficit irrigation in Flame seedless grapevine via Paulsen interstock. Revista Brasileira de Fruticultura 44(1): e-839.
Goldschmidt, E.E.
2014. Plant grafting: New mechanisms, evolutionary implications. Frontiers
in Plant Science 5: 727.
González-Mas, M.C., Rambla, J.L., López-Gresa, M.P., Amparo Blázquez, M.
& Granell, A. 2019. Volatile compounds in citrus essential oils: A
comprehensive review. Frontiers in Plant Science 10: 12.
Gutiérrez, M.A., Chávez, B.H., Gallegos, R.M.Z.
& Borja, S.F. 2022. Characterization of natural extracts obtained by
supercritical fluids (SCF) from Mashua tubers (Tropaeolum tuberosum) grown in Ecuador. International
Journal of Health Sciences 6(S8): 1286-1293.
He, W., Chai, J., Xie, R., Wu, Y., Wang, H.,
Wang, Y., Chen, Q., Wu, Z., Li, M., Lin, Y., Zhang, Y., Luo, Y., Zhang, Y.,
Tang, H. & Wang, X. 2024. The effects of a new citrus rootstock Citrus junos cv. Shuzhen No. 1 on
performances of ten hybrid citrus cultivars. Plants 13(6): 794.
Ho, C.H., Yang, M.H. & Lin, H.L. 2021.
Temperature and different organs create volatile profile differences of edible gynura [Gynura bicolor (roxb. ex willd.) dc]. HortScience 56(8): 954-960.
Johannes, E., Tuwo,
M., Katappanan, N., Henra,
H. & Wirianti, G. 2022. Edible coating berbasis pati ubi kayu Manihot esculenta Crantz dan Jahe Merah Zingiber
officinale var. rubrum memperpanjang umur simpan buah tomat Solanum lycopersicum L. Agrotrop: Journal on Agriculture Science 12(2): 204.
Khalil, N.H. 2023. The effect of organic
fertilization on leaf mineral content of three citrus species. IOP
Conference Series: Earth and Environmental Science 1158(4): 042049.
Killiny, N., Valim,
M.F., Jones, S.E. & Hijaz, F. 2018. Effect of different rootstocks on the
leaf metabolite profile of ‘Sugar Belle’ mandarin hybrid. Plant Signaling
and Behavior 13(3): e1445934.
Kumar, S., Awasthi, O.P., Dubey, A. &
Sharma, R.M. 2019. Effect of different rootstocks on growth, leaf sclerophylly,
and chlorophyll fractions of Kinnow mandarin. Indian
J. Hort. 74(4): 505-509.
Li, Q., Pan, H., Hao, P., Ma, Z., Liang, X.,
Yang, L. & Gao, Y. 2024. Mechanisms underlying the low-temperature
adaptation of 17β-estradiol-degrading bacterial strain Rhodococcus sp. RCBS9: Insights from physiological and transcriptomic analyses. Frontiers
in Microbiology 15: 1465627.
Li, Y., Xin, G., Liu, C., Shi, Q., Yang, F.
& Wei, M. 2020. Effects of red and blue light on leaf anatomy, CO2 assimilation and the photosynthetic electron transport capacity of sweet pepper
(Capsicum annuum L.) seedlings. BMC Plant Biology 20: 318.
Liao, L., Li, Y., Bi, X., Xiong, B., Wang, X.,
Deng, H., Zhang, M., Sun, G., Jin, Z., Huang, Z.
& Wang, Z. 2022. Transcriptome analysis of Harumi tangor fruits:
Insights into interstock-mediated fruit quality. Frontiers
in Plant Science 13: 995913.
Luo, Z., Zhou, W., Xie, T., Xu, W., Shi, C.,
Xiao, Z., Si, Y., Ma, Y., Ren, Q., Di, L. & Shan, J. 2024. The role of
botanical triterpenoids and steroids in bile acid metabolism, transport, and
signaling: Pharmacological and toxicological implications. Acta Pharmaceutica Sinica B 14(8):
3385-3415.
Mahmoud, L.M., Huyck, P.J., Vincent, C.I.,
Gmitter, F.G., Grosser, J.W. & Dutt, M. 2021. Physiological responses and
gene expression patterns in open-pollinated seedlings of a pummelo-mandarin
hybrid rootstock exposed to salt stress and Huanglongbing. Plants 10(7):
1439.
Marín, J., Rincón Barón, E.J. & Montoya-Lerma, J. 2020. Foliar anatomy of
ten genotypes of the plant Manihot esculenta (Euphorbiaceae). Rev. Biol. Trop. (Int. J. Trop. Biol.) 68(3): 919-932.
Negi, J., Munemasa,
S., Song, B., Tadakuma, R., Fujita, M., Azoulay-Shemer,
T., Engineer, C.B., Kusumi, K., Nishida, I.,
Schroeder, J.I. & Iba, K. 2018. Eukaryotic lipid metabolic pathway is
essential for functional chloroplasts and CO2 and light responses in
Arabidopsis guard cells. Proceedings of the National Academy of Sciences of
the United States of America 115(36): 9038-9043.
Noreen, H., Smith, E.N., Farman, M., Claridge,
T.D.W. & McCullagh, J.S.O. 2021. Isolation, separation, identification, and
quantification of bioactive methylated flavone regioisomers by UHPLC-MS/MS. Analytical Science Advances 2(7-8): 364-372.
Oliveira, J.A.A., Bruckner, C.H., da Silva,
D.F.P., dos Santos, C.E.M., dos Santos Soares, W. & Nunes, L.V. 2019.
Performance of interstocks in the plant development
and fruit quality of plum trees. 41(1): 39928.
Rasool, A., Mansoor, S., Bhat, K.M., Hassan,
G.I., Baba, T.R., Alyemeni, M.N., Alsahli,
A.A., El-Serehy, H.A., Paray,
B.A. & Ahmad, P. 2020. Mechanisms underlying graft union formation and
rootstock scion interaction in horticultural plants. Frontiers in Plant
Science 11: 590847.
Raveau, R., Fontaine,
J. & Lounès-Hadj Sahraoui, A. 2020. Essential oils as potential alternative
biocontrol products against plant pathogens and weeds: A review. Foods 9(3): 365.
ReportLinker. 2025. Forecast: Citrus Production in Indonesia. ? (Accessed on August 25, 2025).
Rong, Y., Liao, L., Li, S., Wei, W., Bi, X.,
Sun, G., He, S. & Wang, Z. 2023. Comparative transcriptomic and
physiological analyses reveal key factors for interstocks to improve grafted seedling growth in Tangor. International Journal of
Molecular Sciences 24(7): 6533.
Shen, Y., Zhuang, W., Tu, X., Gao, Z., Xiong,
A., Yu, X., Li, X., Li, F. & Qu, S. 2019. Transcriptomic analysis of interstock-induced dwarfism in Sweet Persimmon (Diospyros
kaki Thunb.). Horticulture Research 6: 51.
Susilo, D.E.H.
2015. Identifikasi nilai konstanta bentuk daun untuk pengukuran luas daun metode panjang kali lebar pada tanaman hortikultura di tanah gambut. Anterior Jurnal 14(2): 139-146.
Taj, T., Sultana, R., Shahin, H.D., Chakraborthy, M. & Gulzar Ahmed, M. 2021. Phytol A
phytoconstituent, its chemistry and pharmacological actions. 8(1): 395-406.
Thiesen, L.A., Pinheiro, M.V.M., Holz, E.,
Werner, A., Eloy, E., Caron, B.O. & Schmidt, D. 2022. Phenotypic plasticity
of Aloysia citrodora:
Anatomical changes to water availability and seasons. Comunicata Scientiae 13: e3590.
Torabi, F., Majd, A. & Enteshari,
S. 2015. The effect of silicon on alleviation of salt stress in borage (Borago
officinalis L.). Soil Science and Plant Nutrition 61(5): 788-798.
Toscano-Morales, R., Xoconostle-Cázares,
B., Martínez-Navarro, A.C. & Ruiz-Medrano, R. 2016. AtTCTP2 mRNA and
protein movement correlates with formation of adventitious roots in tobacco. Plant
Signaling and Behavior 11(3): e1071003
Trend Economy. 2024. Indonesia Imports and Exports of World Citrus Fruit 2012-2023. (Accessed on 25 August 2025).
Vives-Peris, V., López-Climent, M.F.,
Moliner-Sabater, M., Gómez-Cadenas, A. & Pérez-Clemente, R.M. 2023.
Morphological, physiological, and molecular scion traits are determinants for
salt-stress tolerance of grafted citrus plants. Frontiers in Plant Science 14: 1145625.
Wang, T., Xiong, B., Tan, L., Yang, Y., Zhang,
Y., Ma, M., Xu, Y., Liao, L., Sun, G., Liang, D., Xia, H., Zhang, X., Wang, Z.
& Wang, J. 2020. Effects of interstocks on growth
and photosynthetic characteristics in ‘Yuanxiaochun’ Citrus
seedlings. Functional Plant Biology 47(11): 977-987.
Widyaningsih, S., Utami,
S.N.H., Joko, T. & Subandiyah, S. 2017.
Development of disease and growth on six scion/rootstock combinations of citrus
seedlings under Huanglongbing pressure. Journal of Agricultural Science 9(6): 229-238.
(WCO). 2022. Produce Report. World Citrus Organization Announces 2021/22 Statistics. (Accessed on 25 August 2025).
Yao, X.C., Meng, L.F., Zhao, W.L.
& Mao, G.L. 2023. Changes in the morphology traits, anatomical structure of
the leaves, and transcriptome in Lycium barbarum L. under salt stress. Frontiers in Plant
Science 14: 1090366.
Yulianti, F. & Agisimanto,
D. 2023. Leaf physiological responses of the citrus commercial varieties
grafted onto rootstocks. IOP Conference Series: Earth and Environmental
Science 1172: 012017.
Zhang, L., Su, Q.F.,
Wang, L.S., Lv, M.W., Hou, Y.X. & Li, S.S. 2023a.
Linalool: A ubiquitous floral volatile mediating the communication between
plants and insects. Journal of Systematics and Evolution 61(3): 538-549.
Zhang, X., Yang, W., Tahir, M.M., Chen, X., Saudreau, M., Zhang, D. & Costes, E. 2023b.
Contributions of leaf distribution and leaf functions to photosynthesis and
water-use efficiency from leaf to canopy in apple: A comparison of interstocks and cultivars. Frontiers in Plant Science 14: 1117051.
Zheng, L. & Van Labeke,
M.C. 2017. Long-term effects of red- and blue-light emitting diodes on leaf
anatomy and photosynthetic efficiency of three ornamental pot plants. Frontiers
in Plant Science 8: 917.
*Corresponding
author; email: nfdevy@gmail.com
|
|||
|
