Quantitative and qualitative traits analyses in the advance breeding lines of rice

Transcription

1 International Journal of Biosciences IJB ISSN: (Print), (Online) Vol. 6, No. 8, p , 2015 RESEARCH PAPER OPEN ACCESS Quantitative and qualitative traits analyses in the advance breeding lines of rice Seema Ali Khan 1, Azhar Hussain Shah 1*, Fida Muhammad Abbasi 2, Arooj Javed 1, Inayat ur Rahman, Habib Ahmad 2 1 Department of Botany, Hazara University, Mansehra Khyber Pakhtunkhwa, Pakistan 2 Department of Genetics, Hazara University, Mansehra, Khyber Pakhtunkhwa, Pakistan Key words: Qualitative, quantitative, rice, line, traits. Article published on April 25, 2015 Abstract In present study fifteen advance lines of rice (Line-95, Line-168, Line-133, Line-50, Line-79-L, Line-146, Line- 149, Line-2, Line-29, Line-35, Line-54, Line-127, Line-64, Line-86-2, Line-71) were evaluated in the background of JP-5 (control) for Quantitative and Qualitative traits. Mean value for quantitative traits were compared according to least significant difference (LSD) statistical test. Data on morphological parameters (qualitative and quantitative) was collected from productive tillers of every three plants of each variety at suitable phase of growth to check variation. Findings were recorded according to IRRI description morphology and varietals characteristics. The material exhibited considerable inconsistency for qualitative and quantitative characters with few exceptions. Genetic distance based on SDS PAGE analysis ranges from 0 70%. Dendogram was constructed which divided sixteen genotypes in six groups (A F) comprising of 3, 2, 5, 1, 3 and 2 genotypes, respectively. The advance rice lines such as Line-2, Line-29, Line-35, Line-54, Line-127 and Line showed good physical characteristics (grain length, grain width, L/B ratio, chalkiness). While grain quality traits (GT, GC) were excellent in advance Line-95, Line-168, Line-2, Line-35, Line-54, Line-127, Line-64 and Line-71. However, highest yield/plant was recorded in Line-86-2 (162g), Line-79-L (134.9g), Line-71 (116.5g), Line-95 (114.1g), Line-168 (110.1g) and Line-149 (103.7g). Our findings showed great variations in morphological, physical and biochemical traits of advance rice lines and these variations may be exploited to develop high yielding and high quality rice either through breeding or by genetic engineering. * Corresponding Author: DR. Azhar Hussain Shah drahshu@gmail.com 50 Khan et al.

2 Introduction Rice (Oryza sativa L.) belongs to family Poaceae of tribe Oryzeae. The genus Oryza consists of two cultivated species Oryza sativa (Asian species) and Oryza glaberrima (African species) and about 20 wild species (Vaughan et al. 2003; Linscombe, 2006). Of these two species, Oryza sativa is cultivated all over the world whereas Oryza glaberrimais cultivated in West Africa only. Oryza sativa is composed of two major biotypes or sub-species indica and japonica and represent evolutionary adaptation to environmental variables (Maclean et al., 2002) Rice is an economically important crop and feeds nearly half the world s population and accounts for more than 50% of their daily calorie intake (Maclean et al., 2002). It is a source of magnesium, thiamin, niacin, phosphorus, vitamin B6, zinc and copper. Some varieties have iron, potassium and folic acid. White rice is one of the poorest cereals in proteins; some improved varieties however may provide 14g of protein per 100g. A number of energy rich compounds such as carbohydrates, fat, protein and reasonable amount of calcium and riboflavin are found in rice (Juliano et al., 1993). Most of rice consumed directly by humans. Apart from providing a significant proportion of the daily calorie and protein intake for half the world population, Oryza sativa is also a livestock feed that has an array of industrial and non-industrial uses. Depending upon environment, soil, variety and fertilizer regimes the chemical composition of rice grains may vary. It has been predicted that the rate of world population increase by 2030 to feed this population we must increase by 70% (Maclean et al., 2002). World production of rice has risen gradually from about 200 million tons of paddy rice in 1960 to over 678 million tons in 2009, while the average world yield for rice was 4.3 tons per hectare, in 2010 (Shabana, 2012). Although rice production has increased substantially, but it falls short of growing demand due to population increase which is rising at a rate of 2% annually. To feed this increasing population, 35% more rice will be required than the present level of rice production globally (Duwayrie and Nguyen, 1999). Growing human populations coupled with adverse and unpredictable environmental conditions and declining yield have become the greatest threats for production today (Childs, 2004). Among the major yield limiting factors, yield stagnation is main threat to productivities that may be due to several, biotic (diseases, insects & pests) and abiotic (soil, temperature & drought) factors. The loss caused by these factors was estimated to be 200 million tons of world rice production. Low yield due to poor crop management techniques, lack of research and extension system, and limited utilization of productive varieties. Good quality seed is expected to increase yield by 5-20 % and that the extent of this increase is directly proportional to the quality of seed that is being sown (Rickman et al., 2006). But the lack of reliable scientific information regarding the agronomically important qualitative and quantitative traits hinders the production of good quality seeds. Keeping this in view present study was designed to evaluate fifteen advance lines of rice in comparison with JP-5 for economically and agronomically important qualitative and quantitative traits; to assess the variability of advance rice lines at protein level, to identify the reliable and distinguishing characters among advance breeding lines of rice which could be linked with rice quality and to identify intraspecific boundaries between different rice lines. Materials and methods Plant materials Fifteen advance lines (Line-95, Line-168, Line-133, Line-50, Line-79-L, Line-146, JP-5, Line-149, Line-2, Line-29, Line-35, Line-54, Line-127, Line-64, Line- 86-2, Line-71) in the background of JP-5 (control) were grown under similar environmental conditions at National Tea Research Institute (NTRI) Shinkiari, Mansehra, KP, Pakistan during 2012 assess variations agronomically important qualitative and quantitative traits. 51 Khan et al.

3 All morphological, physiological and agronomic traits such as stem length, stem thickness, productive tillers/plant, leaf length, leaf width, panicle length, branches /panicle, yield/plant (g), sterility % age, 1000 grains weight, grain chalkinesss, gelatinization temperature (GT), gel consistency (GC) and aroma in elite rice lines were recorded following IRRI standard descriptor. Proteins Estimation and Expressions For water soluble total protein extraction/separation Sambrook et al. (1989) method was used. Statistical analysis Genetic diversity was measured by using the following formula of Nei and Li, (1979). GD = 1 - dxy/dx + dydxy Where GD is the genetic distance between two genotypes, dxy is the number of common bands in 2 nd genotypes, dx is the total number of bands in genotype 1 st and dy is total number of bands in genotype 2 nd. The bi-variant 1-0 data matrix was also used to construct a dendrogram by using computer program Popgene while for the analysis of other quantitative traits statistix 8.1 trial version software was used. Results Under similar environmental conditions 15 advance lines with JP-5 showed considerable variability for morphological characters including both qualitative and quantitative traits. Tillers/plant It ranged from to Maximum tillers/plant was recorded for Line-168 (36.00) followed by Line- 54 (29.67), Line-29 (26.33). Minimum tillers/plant was recorded for Line-133 (10.33), (Table 1). Table 1. Means for tillers/plant, stem length, stem thickness, ligule length of advance lines of rice. Variety Tillers/plant (no) Stem length (cm) Stem thickness (cm) Ligule Length (mm) Line defgh cd 2.40 d 1.90 a Line a efg 1.60 j 1.70 f Line h def 1.80 i 2.00 c Line efgh ab 2.90 a 1.90 d Line-79-L cdefg bc 2.50 c 1.80 e Line gh g 2.07 f 1.70 f Line-JP bcd a 2.00 g 1.90 d Line fgh def 1.57 j 1.40 i Line efgh cd 2.70 b 2.70 b Line bc efg 1.90 h 1.70 f Line bcdef cd 1.90 h 1.90 d Line ab cde 1.80 i 1.50 h Line bcde fg 1.90 h 1.90 d Line cdef def 2.00 g 2.73 b Line defgh ab 1.40 k 1.57 g Line defgh cd 2.20 e 4.80 a CV% Mean sharing same letters in a column are not significantly different at 5% probability level using LSD test. Stem length It ranged from to Maximum stem length was recorded for JP-5 ( cm) followed by Line-50 (153 cm), Line-86-2 ( cm). Minimum stem length was recorded for Line-146 (80.73 cm), (Table 1). Stem thickness 52 Khan et al.

4 It ranged from 1.40 to Maximum stem thickness was recorded for Line-50 (2.90 cm) followed by Line- 2 (2.70 cm), Line-79-L (2.50 cm). Minimum stem thickness was recorded for Line-86-2 (1.40 cm), (Table 1). Ligule length It ranged from 1.40 to Maximum ligule length was recorded for Line-71 (4.80 mm) followed by Line- 64 (2.73 mm), Line-2 (2.70 mm). Minimum ligule length was recorded for Line-149 (1.40 mm), (Table 1) Leaf length It ranged from to Maximum leaf length was recorded for Line-64 ( cm) followed by Line-71 (87.10 cm), Line-79-L (82.30 cm). Minimum leaf length was recorded for Line-29 (46.90 cm), (Table 2). Table 2. Means for leaf length, leaf length of blade, leaf width, flag leaf length, flag leaf width of advance lines of rice. Variety Leaf Length (cm) Leaf Length of Blade (cm) Leaf Width (cm) Flag Leaf Length (cm) Flag Leaf Width (cm) Line i d 1.40 a j 1.90 b Line l h 1.20 ab i 1.40 f Line g j 1.07 bc o 1.40 f Line c c 1.30 ab p 1.80 c Line-79-L c e 1.10 bc b 1.80 c Line j l 0.90 c d 1.90 b JP b d 1.30 ab f 1.60 d Line k k 1.27 ab e 2.00 a Line e h 1.33 ab g 1.40 f Line n m 1.10 bc n 1.30 g Line h g 1.10 bc h 1.40 f Line i i 1.43 a k 1.40 f Line m m 1.27 ab l 1.30 g Line a a 1.23 ab m 1.50 e Line f f 1.30 ab c 1.60 d Line b b 1.20 ab a 1.80 b CV% Leaf length of blade It ranged from to Maximum leaf length of blade was recorded for Line-64 (79.00 cm) followed by Line-71 (64.63 cm), Line-50 (60 cm). Minimum value was recorded for Line-29 (26.60 cm), (Table 2). Leaf width It ranged from 0.90 to Maximum leaf width was recorded for Line-54 (1.43 cm) followed by Line-95 (1.40 cm), Line-2 (1.33 cm). Minimum leaf width was recorded for Line-146 (0.90 cm), (Table 2). Flag Leaf Length It ranged from to Maximum flag leaf length was recorded for Line-71 (91.17 cm) followed by Line-79-L (76.80 cm), Line-86-2 (74.50 cm). Minimum value was recorded for Line-50 (40.10 cm), (Table 2). Flag Leaf Width It ranged from 1.30 to Maximum flag leaf width was recorded for Line-149 (2.00 cm) followed by Line-95 (1.90 cm), Line-146 (1.90 cm). Minimum flag leaf width was recorded for Line-127 (1.30 cm) and Line-29 (1.30 cm), (Table 2). Panicle length It ranged from to Maximum panicle length was recorded for Line-86-2 (35.46 cm) followed by Line-71 (31.67 cm), Line-95 (31.20 cm). Minimum panicle length was recorded for Line-127 (21.40 cm), (Table 3). 53 Khan et al.

5 Table 3. Means for panicle length, primary branches, secondary branches, filled spikelets, unfilled spikelets of advance lines of rice. Variety Panicle length(cm) Primary Branches(no) Secondary Branches(no) Filled Spikelet(no) Unfilled Spikelet (no) Line b a ab bc ab Line cd h ef defg cde Line ef efg f g e Line b ab abc bcd ab Line-79-L b bc bc cdef ab Line bc e bcd efd a JP de ef cde bcde bc Line bc ab a bc ab Line bc cd ab bc bc Line ef fgh def g de Line b ef cde bcde cde Line ef h f g e Line f gh def g de Line def efgh ef fg cde Line a de abc b ab Line b bc a a bcd CV% Mean sharing same letters in a column are not significantly different at 5% probability level using LSD test. Primary branches It ranged from to Maximum primary branches was recorded for Line-95 (20.10 cm) followed by Line-50 (18.99 cm), Line-79-L (17.55 cm). Minimum primary branches was recorded for Line-54 (10.10 cm) and Line-54 (10.10 cm), (Table 3). Secondary branches It ranged from to Maximum secondary branches was recorded for Line-149 (66.11 cm) followed by Line-2 (59.44 cm), Line-71 (64.77 cm). Minimum secondary branches was recorded for Line- 133 (19.67 cm), (Table 3). Filled spikelets It ranged from to Maximum filled spikelets was recorded for Line-71 ( cm) followed by Line-86-2 ( cm), Line-95 ( cm). Minimum filled spikelets was recorded for Line- 54 ( cm), (Table 3). Unfilled spikelets It ranged from to Maximum unfilled spikelets was recorded for Line-146 (71.66 cm) followed by Line-149 (60.89 cm), Line-79-L (54.44 cm). Minimum valuewas noted for Line-133 (13.33 cm), (Table 3). Physico-chemical properties of advance lines of rice Physico-chemical properties directly linked to cooking and eating feature of rice that classify the accepted rice grain quality, such as following: Qualitative Traits Grain size it is one of the key character related to the cosumers preference. JP-5, Line-95, Line-146, Line-149, Line- 35 and Line-127 have short grain while Line-64 has medium size grain and all other lines have long grain, (Table 4). Grain shape JP-5, Line-95, Line-146, Line-149 and Line-127 have medium shape while Line-168, Line-133, Line-50, Line-79-L, Line-71, Line-29, Line-35, Line-54, Line 64, Line-86-2 and Line-2 have slender shape grain, (Table 4). Gelatinization temperature JP-5, Line-95, Line-168, Line-79-L, Line-146, Line- 149, Line-35, Line-127 and Line-86-2 have intermediate gelatinization temperature while Line- 133, Line-50, Line-54, Line-71, Line-64 and Line-2 have low and Line-29 have high gelatinization temperature, (Table 4). 54 Khan et al.

6 Table 4. Evaluation of advance lines of rice for qualitative traits of grain. Variety Size Shape GT Aroma GC Line-95 Short Medium Intermediate NS Soft Line-168 Long Slender Intermediate MS Soft Line-133 Long Slender Low NS Hard Line-50 Long Slender Low SS Medium Line-79-L Long Slender Intermediate MS Medium Line-146 Short Medium Intermediate SS Medium JP-5 Short Medium Intermediate NS Soft Line-149 Short Medium Intermediate MS Medium Line-2 Long Slender Low NS Soft Line-29 Long Slender High NS Soft Line-35 Short Slender Intermediate SS Soft Line-54 Long Slender Low MS Soft Line-127 Short Medium Intermediate NS Soft Line-64 Medium Slender Low NS Soft Line-86-2 Long Slender Intermediate SS Medium Line-71 Long Slender Low NS Soft Aroma JP-5, Line-95, Line-133, Line-71, Line-29, Line-127, Line-64 and Line-2 have no aroma while Line-168, Line-79-L, Line-149, Line-54 have mild aroma and Line-50, Line-146, Line-35 and Line have very strong aroma (Table 4). Gel consistency JP-5, Line-95, Line-168, Line-71, Line-29, Line-35, Line-54, Line-127, Line-64 and Line-2 have soft gel consistency and all other advance lines of rice have medium except Line-133 which have hard gel consistency, (Table 4). Quantitative Traits Grain length It ranged from 5.02 to Maximum grain length was recorded for Line-133 (7.30 mm) followed by Line-79-L (6.77 mm), Line-149 (5.07 mm). Minimum grain length was recorded for JP-5 (5.02 mm), (Table 5). Table 5. Means for grain length, grain width, L/B ratio and kernel chalkiness% of advance lines of rice. Variety Grain length (mm) Grain width (mm) L/B ratio Kernal Chalkiness % Line d 1.77 ab 2.87 d b Line b 1.75 b 3.83 b d Line a 1.65 g 4.41 a a Line b 1.67 efg 4.02 b 0.00 f Line-79-L 6.77 b 1.67 fg 4.06 b e Line d 1.77 ab 2.87 d cd JP d 1.78 a 2.81 d 0.00 f Line d 1.76 ab 2.88 d bc Line b 1.67 efg 4.01 b 0.00 f Line b 1.70 cd 3.93 b 0.00 f Line d 1.69 cde 3.02 d 0.00 f Line b 1.70 cde 3.93 b 0.00 f Line d 1.71 c 2.97 d 0.00 f Line c 1.76 ab 3.50 c e Line b 1.68 defg 3.98 b 0.00 f Line b 1.69 cdef 3.97 b e CV% Mean sharing same letters in a column are not significantly different at 5% probability level using LSD test. 55 Khan et al.

7 Grain width It ranged from 1.65 to Maximum grain width was recorded for JP-5 (1.78 mm) followed by Line-95 (1.77 mm), Line-146 (1.77 mm). Minimum grain width was recorded for Line-133 (1.65 mm), (Table 5). L/B ratio It ranged from 2.81 to Maximum L/B ratio was recorded for Line-133 (4.41) followed by Line-79-L (4.06), Line-50 (4.02). Minimum L/B ratio was recorded for JP-5 (2.81), (Table 5). Kernal chalkiness It ranged from to Maximum kernel chalkiness was recorded for Line-133 (83.33%) followed by Line-95 (61.10%), Line-149 (55.55%). Minimum kernel chalkiness was recorded for Line-64 (19.04%), (Table 5). Yield/plant (g) It ranged from 45.8 to 162. Maximum yield/plant was recorded for Line-86-2 (162g) and minimum for 133 (45.8g), Table (6). Table 6. Evaluation of advance lines of rice for quantitative traits. Variety No. nodes /stem Yield/plant (g) Sterility% 1000grain wt(g) GC, (mm) Line Line Line Line Line-79-L Line JP Line Line Line Line Line (above) Line Line Line Line (above) Sterility% It ranged from to Maximum sterility was recorded for Line-146 (30.88%) followed by Line-79- L (26.82%), Line-50 (25.09%). Minimum was recorded for Line-71 (10.44%), (Table 6). 1000grain weight (g) It is very important feature for the estimation of yield productivity. Maximum grain weight was recorded for Line-133 and Line-146 (33.2g) while minimum recorded for Line-2 (16.8g), (Table 6). Gel consistency Maximum GC was recorded in Line-71, Line-54 and Line-29 (100) while minimum GC value was recorded for Line-133 (34), (Table 6). Assessment of Genetic Diversity through SDS-PAGE Genetic diversity based on seed storage proteins in the rice genotypes is presented in Table 7. Only clearly score able bands were included in the study. Every individual band was considered as a single allele. A total of 7 bands were scored in the sixteen germplasm used during present study. Locus (Bands) numbers 6 and 7 were present in all the sixteen genotypes. Genetic distance observed among the sixteen germplasm was ranged from 0 70%. Minimum GD (0%) were recorded among 20 comparisons while for only one comparison maximum genetic distance (70%) was observed. 56 Khan et al.

8 Genetic distances for 0, 25, 44, 16, 12 and 3 comparisons were recorded as 10, 20, 30, 40, 50, and 60 %, respectively. Dendrogram based on SDS-PAGE analysis grouped all the 16 genotypes in six groups A, B, C, D, E and F Fig 1. Group C comprised of 5 genotypes while groups A, B, D, E and F, comprised 3, 2, 1, 3 and 2 genotypes, respectively. Line-95 and Line-71 were found most distantly allied to each other and hence in future breeding programs, these two lines (line-95 and line-71) should be crossed to create maximum genetic variability in rice breeding population. Table 7. Genetic distances among 16 advance lines used during present study = Line-95, 2= Line-168, 3= Line-133, 4= Line-50, 5= Line-79-L, 6=Line-146, 7= Line-JP-5,8= Line-149, 9=Line- 2, 10= Line-29, 11= Line-35, 12= Line-54, 13= Line-127, 14= Line-64, 15= Line-86-2, 16= Line-71. Discussion To enhance rice production to meet consumption requirements and to improve the grain quality more efforts are required. Because consumer preference is influenced by rice grain size, shape, color, aroma, gelatinizing temperature and chalkiness rice breeders must take all these parameters into consideration when developing a new variety (Vidal et al., 2007). Fifteen advance lines of rice and JP-5 were studied for different qualitative and quantitative traits to assess the genetic variability. The basic statistical data (mean, range, variance, standard deviation and standard error) for each of the quantitative traits measured for all the advance lines. The materials showed considerable variability for both qualitative and quantitative traits. These results are in agreement with Zafar et al., (2006a) who evaluated Landrace genotypes of rice collected from various parts of Pakistan for seven quantitative and eight qualitative characters and a significant amount of genetic variation was displayed for most of the traits examined. Our results revealed that morphology varies among the advance lines of rice and the rest of the traits positively correlated to each other. For the improvement of grain quality and potential yield it is essential to identify the morphological traits that could contribute for the best quality and increasing yield potential of rice. The rice grain quality can be determined by both physical and chemical characteristics. These characteristics may be genetic or obtained. The marketing of rice mainly depends on the physical dimensions (grain size, shape) and weight which are 57 Khan et al.

9 of prime importance (Rice Quality Workshop, 2003). Our results revealed that physical traits of grains varies among the advance lines of rice and these results are in conformity with Taylor and Parker, (2002) who reported that rice grains vary in length and are of great importance to consumers, producers and breeders because most of the rice produced in the world is consumed as a entire grain. The results of physical parameters indicated that the highest grain length (7.30 mm) was found in advance Line-133 and the lowest (5.02 mm) was found in JP-5 which categorized the size of Line-133 and JP-5 as long and short respectively. These findings are in conformity with Rickman el al., (2006) they also used the length of the rice milled kernel for determination of its size. While the highest grain width (1.7 mm) showed by JP-5 and the lowest grain width was found in Line- 133 (1.6 mm). Coefficients Fig. 1. Dendrogram constructed for sixteen advance lines of rice using protein profiles obtained by SDS- PAGE. The ratio of length to width is an important feature to determine the shape of the variety. In present study the lines were categorized into two types, slender (11 lines) and medium (5 lines) based on their length to width ratio. The L/B ratio was also use by Rickman et al., (2006) for shape to classified milled grain. Generally, consumers in the tropics and sub tropics select medium and slender grains and in temperate areas, short and bold grains are selected (Khush, 2001). Grain weight is an important yield component because it gives the information about the size and density of the rice grains. The density of different rice grains effect the cooking and eating qualities. So grain weight should be uniform because it determines the quality of rice grains (Rice CRC, 2003). The highest 1000 grain weight was recorded in two varieties Line- 133 (33.2g, long) and Line-146 (33.2g, medium). These results differ from IRRI (2009) which reported that medium or bold grains are heavier in weight than longer grains. The rice varieties having minimum amount of chalkiness is consider as good quality grains in comparison with chalky once which decrease the rice grain quality. In present investigation the highest chalkiness% was recorded in Line-133 (83.33) while the chalkiness% recorded for Line-95, Line-168, Line- 79-L, Line-146, Line-149 and Line-71 were ranged from These results are in conformity with Shilpa and Krishnan, (2010) who reported that in high yielding varieties the chalkiness found to be less and ranged from %. While the Line-2, Line-35, Line-54, Line-29, Line-127, Line-86-2, Line- 50 and JP-5 were not chalky, hence acceptable for cosumers with better returns. And these findings are in conformity with Cheng et al., (2005) who reported that the chalky rice grains reduce the palatability of cooked products, therefore the presence of more than 20% chalkiness in rice kernels is not acceptable in world markets. Gelatinization temperature is a physical property of starch grain that predicts the time and energy required for cooking rice. In present findings nine lines were intermediate (partially degraded), six were low and the only one variety (29) was recorded for high GT. These results are in favour of Kang et al., (2006) who reported that intermediate to low GT varieties take less time to cook than having high GT varieties and are preferred by some consumers. And 58 Khan et al.

10 also same results were given by Shabana (2012) who reported positive relation between GT and volume expansion of the cooked grain, while negative relation between GT and kernel elongation. The gel consistency is responsible for softness (Sabouri, 2009). Our findings revealed that ten soft advance lines of rice (Line-2, Line-95, Line-133, Line- 29, Line-35, Line-54, Line-127, Line-64, Line-71 along with check variety JP-5) would be preferred in the East Asian market, as they prefer sticky cooked rice. These findings are in conformity with Tang et al., (1991) who reported that rice with soft gel consistency cook more tenderly and remain soft even upon cooling. Therefore, preferred by consumers due to their tender texture. While the only one Line-133 was hard. Aroma is another important trait in rice and the aromatic rice has high demand in the market. Pakistan is primarily known for its aromatic rice as it accounts for 2% of exports and replacing India as the only source of aromatic rice (Basmati) in the market. In our findings Line-50, Line-146, Line-35 and Line were strongly scented while Line-168, Line-149, Line-54 and Line-79-L were mild scented. These findings are in conformity with Shilpa and Krishnan, (2010) who studied the physicochemical and cooking characteristics for 22 traditionally cultivated rice varieties from Goa, in comparison with three high yielding rice lines. Among the rice varieties examined for aroma, Annapurna, Barik Kudi, Salsi, TamdeJyoti, IR8, Jyoti and Jaya showed the presence of mild aroma. The fractionation of seed protein by polyacrylamide gel electrophoresis (PAGE) is used as an supplementary tool for assessing species relationship (Sammour, 1989). In the present study considerable level of genetic variation was observed in the fifteen advance lines of rice along with JP-5, germplasm. While relatively lower level of genetic distances (GD = 0%-70%) were examined in the advance lines. Dendogram was constructed which divided sixteen genotypes in six groups (A F) comprising 3, 2, 5, 1, 3 and 2 genotypes, respectively. Line-95 and Line-71 were found most distantly related to each other. Similar results were given by Asghar et al., (2004) he analysed 20 accessions of rice for total seed protein through SDS-PAGE for the purpose of interspecific and intraspecific variation. A considerable variation in protein banding pattern of these rice genotypes was observed. Based on the results of present study, it can be concluded that the rice lines showed great diversity in their qualitative and quantitative traits. Physical (Grain width, grain length, L/B ratio, chalkiness) and chemical (GT, GC, aroma) traits varied among the advance rice lines and are closely associated with grain quality. The protein bands which are common in JP-5 (Japonica rice) and all advance lines of rice might be involved for the development of Japonica nature of rice. These advance lines have different quality characters for different areas of consumers, therefore valuable in local as well as international market, as in international market long grain aromatic rice like Line-50 and Line-86-2 is highly popular and the most expensive. Data on physical characteristics could be used to select promising and adaptable lines that are preferred by consumers, as for tropics and sub-tropics areas advance Line-168, Line-133, Line-50, Line-79-L, Line-29, Line-86-2, Line-71, Line-54 and Line-2 are recommended. Genotypes 95 and 71 were found most distantly related to each other and hence it is recommended that in future breeding programs, these two genotypes (Line-95 and Line-71) should be crossed to get most promising lines of rice. The advance Line- 50, Line-146, Line-35 and Line-86-2 would be preferred by consumers in the world market due to their strong aroma and keep up Pakistan as the only source of aromatic rice. Today there are only a few commercially available chalk-free rice varieties. Therefore, the chalky-free rice Line-2, Line-35, Line- 54, Line-29, Line-127, Line-86-2 and Line-50 will help to Pakistan s economy. High yielding rice lines such as Line-86-2, Line-79-L, Line-71, Line-95, Line- 168 and Line-149 could be used in biotechnological research for more improvement. 59 Khan et al.

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