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African Crop Science Journal, Vol. 27, No. 2, pp. 253 - 265 ISSN 1021-9730/2019 $4.00 Printed in Uganda. All rights reserved © 2019, African Crop Science Society African Crop Science Journal by African Crop Science Society is licensed under a Creative Commons Attribution 3.0 Uganda License. Based on a work at www.ajol.info/ and www.bioline.org.br/cs DOI: https://dx.doi.org/10.4314/acsj.v27i2.10 KARYOTYPE STUDIES ON TEN GENOTYPES OF ONION GROWN IN JOS PLATEAU, NIGERIA S.A. SIRAJO and O.A.T. NAMO1 Department of Botany, Faculty of Science, P.M.B. 146, Federal University Lafia, Nasarawa State, Nigeria 1Cytogenetics and Plant Breeding Unit, Department of Plant Science and Technology, University of Jos, P.M.B. 2084, Jos, Plateau State, Nigeria Corresponding author: [email protected] (Received 2 September 2018; accepted 7 May 2019) ABSTRACT The large number of inter- and intra-specific differences in the genus Allium has necessitated karyotype analysis of the cultivated species, in order to describe patterns and directions of chromosomal evolution, which affects morphological differences. In this study, ten genotypes of onion (Allium cepa L.) (Ares, Violet de Galmi, Red Creole, “Wase”, “Dan Zaria”, “Dan Garko”, “Dan Giyawa”, “Bahaushe”, “Bakana” and “Yar Aleiro”) were used for karyotype analysis in the Cytogenetics Laboratory of the University of Jos, Nigeria. Fresh root tips (1-1.5 cm long) obtained from each genotype were prepared and stained with aceto-orcein for 12 minutes. Each stained root tip was placed in a drop of water on a slide, covered with a coverslip and squashed. Each prepared slide was mounted on the microscope to observe the different stages of mitotic division. The results showed that the arm ratio, centromeric index, coefficient of variation, total form and disparity index varied with genotypes. Metacentric, sub-metacentric and sub-telocentric chromosomes were also observed. Based on a dendrogram of the phylogenetic relationships, the genotypes were grouped into four clusters. Cluster I comprised of genotypes Ares, “Wase” and “Bakana”. Cluster II comprised of genotypes “Dan Garko”, Violet de Galmi and “Dan Giyawa”. Cluster III consisted of the genotypes Red Creole, “Bahaushe” and “Dan Zaria”, while Cluster IV comprised of genotype “Yar Aliero”. The principal component analysis revealed that the centromere position and variation in complement length accounted for 92.71% of the total variations among the genotypes. These attributes could be responsible for the differences in fresh bulb size and dry matter yield of onion. Key Words: Allium cepa, chromosome, metacentric, phylogenetic RÉSUMÉ Le grand nombre de différences inter et intraspécifiques dans le genre Allium a nécessité une analyse caryotypique des espèces cultivées pour décrire les modèles et les directions de l’évolution chromosomique, ce qui affecte les différences morphologiques. Dans cette étude, dix génotypes 254 S.A. SIRAJO and O.A.T. NAMO d’oignon (Allium cepa L.) (Ares, Violet de Galmi, Créole rouge, «Wase», «Dan Zaria», «Dan Garko», «Dan Giyawa», «Bahaushe», «Bakana» et «Yar Aleiro») ont été utilisés pour l’analyse du caryotype au laboratoire de cytogénétique de l’Université de Jos, au Nigéria. Des extrémités de racines fraîches (de 1 à 1,5 cm de long) obtenues à partir de chaque génotype ont été préparées et tachées à l’acéto- orcéine pendant 12 minutes. Chaque extrémité de racine tachée a été placée dans une goutte d’eau sur une lame, recouverte d’une lamelle couvre-objet et écrasée. Chaque lame préparée a été placée sur le microscope pour observer les différentes étapes de la division mitotique. Les résultats ont montré que le ratio des bras, l’indice centromérique, le coefficient de variation, la forme totale et l’indice de disparité variaient selon les génotypes. Des chromosomes métacentriques, sous-métacentriques et sub-télocentriques ont été observés. Le dendrogramme des relations phylogénétiques a montré que les génotypes étaient regroupés en quatre groupes. Le groupe I comprend les génotypes Ares, «Wase» et «Bakana». Le groupe II comprend les génotypes «Dan Garko», Violet de Galmi et «Dan Giyawa». Le groupe III était constitué des génotypes Red Creole, «Bahaushe» et «Dan Zaria», tandis que le groupe IV était constitué du génotype «Yar Aliero». L’analyse en composantes principales a révélé que la position du centromère et la variation de la longueur du complément représentaient 92,71% du total des variations parmi les génotypes. Ces caractéristiques pourraient être responsables des différences de taille des bulbes frais et de rendement en matière sèche de l’oignon. Mots Clés: Allium cepa, chromosome, métacentrique, phylogénétique INTRODUCTION related to morphological differences in some cases. A general phenomenon observed in both Onion (Allium cepa L.) is grown for a variety the plant and animal kingdoms is that of purposes, ranging from kitchen to factory- chromosome variation has accompanied made products and for dehydration. It is evolutionary divergence in the taxa studied valued for its distinct pungent flavour, and is (Paknia and Karimzadeh, 2010). an essential ingredient in cuisine (Singh et al., Chromosome identification is essential for 2013). Onion is also used world wide to treat biotechnological studies, including genome diseases such as blood sugar, rheumatism, analysis, somatic hybridisation and ploidy cancer, digestive disorders and prolonged manipulation (Paknia and Karimzadeh, 2010). cough (Singh et al., 2013). Chromosomal aberrations are common in Onion breeding and seed-production varieties of the same plant species, due to programmes world wide have generated multiple translocations, which sometimes short-day, day-neutral and long-day cultivars. involve eight or even ten chromosomes as in Today, almost 90% of the cultivars used in the case of garlic. different parts of the world come from local The cytogenetic characteristics of Allium breeding programmes (Paknia and species have been observed to vary with the Karimzadeh, 2010). geographic location of the plants. This has The chromosomes of Allium have been necessitated the several studies of Allium studied for decades (Fritsch et al., 2001; Cui species, including Allium cepa (Awe and et al., 2008) for their diversity in size, Akpan, 2017). New species are still being structure and number. Variations in karyotypes documented from different parts of the world, are common, between and within species (Cui hence the need for continuous cytogenetic et al., 2008). Vijayavalli and Mathew (1990) studies on them (Oyuntsetsegel et al., 2013; reported the existence of intraspecific Tzanoudakis and Trigas, 2015; Awe and Akpan, polyploidy within several species of Allium. 2017). This paper reports the results of the They also asserted that chromosomal karyotype studies on ten genotypes of onion differences in terms of number or size are cultivated in different parts of Nigeria. Karyotype studies on ten genotypes of onion 255 MATERIALS AND METHODS of forceps. It was allowed to remain in the stain for 10-15 minutes, to pick up the stain. Onion seedlings of the genotypes used in this Triplicates of well-spread metaphase plates study were raised in the green house of the in terms of clarity and organisation, from Federal College of Forestry, Jos, Plateau State. different individuals, were processed and Topsoil, sharp sand and manure (cow dung) analysed for each genotype. The best in the ratio of 2:1:1 were thoroughly mixed metaphase plates were photographed using an and set in ten plastic trays, measuring 70 cm external camera (Celestron digital microscope in diameter, 23 cm in height and 23 cm in imager 2.0) attached to the BX50 Olympus length. microscope, and scanned at 100-resolution. The karyotype study was conducted in the All measurements were recorded using the Cytogenetics Laboratory of the University of software Image J (Abramoff et al., 2004). For Jos, in north-central Nigeria. Nine out of the numerical characterisation, the following ten onion genotypes (Ares, Violet de Galmi, parameters were assessed: Red Creole, “Dan Zaria”, “Dan Giyawa”, “Dan Garko”, “Bahaushe”, “Bakana” and “Yar Chromosome Arm Ratio (AR). The arm Aleiro”); were sourced from the Centre for ratio (AR) of each pair was calculated using Pastoral and Agricultural Research, Usmanu the formula: Danfodiyo University, Sokoto, Nigeria; while a farmers’ variety, ‘Wase’, was obtained from Wase Local Government Area of Plateau State AR = in Nigeria. The agronomic characteristics of the genotypes used in this study are shown in Where: Table 1. L = Long arm length ; and S = Short arm Karyotype study. Fresh root tips of 1-1.5 length cm long were cut from rapidly growing seedlings (3-4 weeks after sowing). The root Total chromosome length (TL). The total tips were pre-treated with 0.5 g colchicine, chromosome length (TL) of each pair was dissolved in 100 ml of distilled water, at room calculated using the formula: temperature for 3 hr. The roots were washed with distilled water for 5 min, at room TL = L+S temperature; after which they were fixed in Carnoy’s fixative (glacial acetic acid: ethanol, Where: 3:1) overnight at room temperature (25 oC). After thorough washing with distilled water, L = Long arm length; and S = Short arm length 1N HCl was added using a dropper. The root tips were allowed to stand in 1N HCl for 10 Relative value (r- value). The ratio of the minutes to hydrolyse the cells. The HCl was shortest to the longest chromosome pair (r- removed from the plastic Petri dish, using the value) was calculated using the formula: dropper. The Petri dish was then refilled with distilled water twice. A microscope slide was placed on a paper towel. Three drops of aceto- r -value = orcein stain were placed in the centre of the slide. The root tip was transferred from the Where: Petri dish to the stain in the slide using a pair 256 S.A. SIRAJO and O.A.T. NAMO TABLE 1.