Investigation of genetic divergence in local sunflower hybrids and inbred lines by applying morphological markers

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Introduction Pakistan has always been challenging, constant scarcity of edible oil and large amount of edible oil provisions are met through import escalating at the rate of 12.5% per annum constantly. Local production meets only 30-35% of edible oil requirements. The gap between consumption and production can be avoided by developing hybrids with high edible oil percentages, early maturity and disease resistance. Presently, sunflower becomes a significant crash crop of country and its area has been increased up to 300614 hectares with the production of 404391 tones (Anonymous 2012). Sunflower (Helianthus annuus L.) is very imperative member of Asteraceae family originated from temperate and tropical regions cultivated widely and more drought tolerant than any other grain crop. It was introduced in 1960s and ranks second after cotton in area and production in Pakistan (Arshad et al., 2012). All over the country crop is grown with 100 percent imported hybrid seed. The imported seed is much expensive and boost up the cost of production. Most of growers cannot afford to purchase hybrid seed every year, because its yield reduced up to 50 % in next generations and also have acclimatization issues in local climatic conditions. Therefore; there is big demand of time to develop local hybrids with higher yield potential and better adoptability (Muhammad et al., 2012). Yield is the combination of numerous organized aspects which are exceptionally influenced by environmental instabilities. It is highly recommended to explore configuration of yield via breeding approaches. The investigation of diverse traits and their association is significant for direct and indirect genetic advancement of crop. The knowledge of the genetic associations among lines can be applied to exploit maximum potential of crop predominantly functional in developing crosses and to identify the particular heterotic fractions, and in accurate estimation with regard to plant varietal conservation. Study of genetic diversity is the practice by which variant individuals or groups of individuals or populations are recognized. (Mudassar et al., 2013). Principle component analysis (PCA) is an appropriate technique; cluster analysis principal component analysis and path coefficient analysis are contemporary mainly very popular advances applied by sunflower researchers for assessment of genetic diversity and for scrutinizing genetic traits and revealed significant consistency regarding conventional breeding procedures. (Mohammadi and Prasanna 2003; Shankar et al., 2006; Arshad et al., 2007; Hidyatullah et al., 2008; Rehman et al., 2013; Ghaffoor 2009). Keeping in view the significance of technique the current investigation was designed with the target to approximate the genetic diversity and character association among sunflower lines and hybrids by applying cluster and principle component analysis. Extensive manuscripts pertaining to genetic diversity and character association in sunflower has been produced earlier by other researchers (Maryyum et al., 2011; Presotto et al., 2009; Darvishzadeh et al., 2011; Amjad et al., 2013). Material and methods The study was conducted in National Agriculture Research centre (NARC), Islamabad, Pakistan during autumn 2011. 10-CMS (Cytoplasmic male sterile lines), 5-Restorers lines along with 17 hybrids were evaluated to explored the genetic diversity regarding numerous agronomic traits. The experimental material CMSHA-03, CMSHA-10, CMSHA-11, CMSHA-18, CMSHA-24 CMSHA-25, CMSHA-54, CMSHA-56, CMSHA-99, CMSHA-103, RHP-73, RHP-42, RHP-46, RHP-73, RHP-77, SMH-1107, SMH-106, SMH-1109, SMH-1030, SMH-1018, SMH- 1007, SMH-1019, SMH-1028, SMH-0933, SMH-1105, SMH-0924, SMH-0920, SMH-0951, SMH-0926, SMH-0945 and SMH-0925 were the NARC (National Agricultural Research Centre), Islamabad locally developed lines and hybrids two imported check hybrids i.e. Hysun-33 (Australia) and NKS-278 (USA) were collected from ICI, Pak seed Pvt. Ltd and Syngenta, Pak seed Ltd. Lahore. The hybrids were replicated using Randomized Complete Block Design (RCBD). Each investigational division holds 5m long Ayaz et al. Page 155
4 rows with row to row spacing of 75cm and plant-toplant 25cm respectively. 3-4 seeds to maintain optimum population per plot were sown on ridges to a depth of 2-3 cm. All other suggested practices were adopted during whole growth period. Optimum fertilizers (120 Kg ha-1 Nitrogen, 60 Kgha-1 Phosphorus) were applied following seed bed preparation and flowering. At maturity ten plants were picked erratically from each division in the central two rows. Observations were carried out regarding: 1) Flower initiation Days (FID) Flower initiation days were calculated from the date of cultivation to 50 percent blooming of flower buds per plot. 2) Full Flowering Days (FFD) The days to full flowering were calculated from the date of cultivation to 90 percent of blooming. 3) Full development Days (FDD) Days to full development were estimated from the date of planting following back of the disk turn yellow and bracts were turn brownish in color. 4) Leaves per plant Leaves of 10 randomly selected plants were counted after attaining of maximum leaves. 5) Height of plant (cm) At full development of crop 10 plants were selected randomly and height was recorded from ground level to the point of attachment of disk with the stem. 6) Disk length (cm) Length of disk of randomly selected plants was measured from one edge of the disc to the other. 7) Thickness of stem (mm) The random plants per plot were selected for recoding thickness of stem from bottom, middle and top of the plant. 8) Weight of 1000 achenes (g) Weight of 1000 achenes was recorded in grams on electric balance from achenes of the selected plants after threshing them manually. 9) Achenes yield (kg/ha). The yield data was calculated at eight percent moisture level by applying formula: Achenes yield at 8% = plot achenes yield at harvest x moisture factor Dry wt. of achenes sample (with paper bag) Moisture factor = Fresh wt. of achenes sample (with paper bag) 10) Oil percentage (%) Oil percentage was determined by using nuclear magnetic resonance (NMR). (Bruker Mini-Spec 10). Statistical analysis Data collected from experiment was analyzed statistically to calculate independently descriptive statistics using computer software MSTAT-C mean association among treatments were estimated using least significant difference (LSD) test at P=0.05 for yield and yield components. The cluster analysis was conducted for presenting genotypes assortment by constructing diversity tree with the help of computer software ‘STATISTICA’ for windows by applying WARD’s method (Sneath and Shokal 1973) mathematical analysis and simple statistics was calculated by using principal component analysis by utilizing computer software 'Past' for Windows. Result and discussion The Range, mean values, coefficient of variation and least significant differences regarding morphological characters are presented in (Table. 1, Table. 2, Table. 3 and Table. 4). The CMS and restorer lines were evaluated only for few characters during initial developmental phases. Flowering and Maturity Traits The highly significant differences were observed among sunflower hybrids and inbred lines at (P
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Investigation of genetic divergence in local sunflower hybrids and inbred lines by applying morphological markers

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