Field Crops Research 101 (2007) 104116 www.elsevier.

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Genetic variability and interrelationship among various morphological and quality traits in quinoa (Chenopodium quinoa Willd.)
Atul Bhargava *, Sudhir Shukla, Deepak Ohri
Division of Genetics and Plant Breeding, National Botanical Research Institute, Lucknow 226001, India Received 29 September 2006; accepted 10 October 2006

Abstract Evaluation of seed yield, morphological variability and nutritional quality of 27 germplasm lines of Chenopodium quinoa and 2 lines of C. berlandieri subsp. nuttalliae was carried out in subtropical North Indian conditions over a 2-year period. Seed yield ranged from 0.32 to 9.83 t/ha, higher yields being shown by four Chilean, two US, one Argentinian and one Bolivian line. Two lines of C. berlandieri subsp. nuttalliae exhibited high values for most of the morphological traits but were low yielding. Seed protein among various lines ranged from 12.55 to 21.02% with an average of 16.22 0.47%. Seed carotenoid was in the range of 1.695.52 mg/kg, while leaf carotenoid was much higher and ranged from 230.23 to 669.57 mg/kg. Genetic gain as percent of mean was highest for dry weight/plant, followed by seed yield and inorescence length. All morphological traits except days to owering, days to maturity and inorescence length exhibited signicant positive association with seed yield. The association of leaf carotenoid with total chlorophyll and seed carotenoid was positive and highly signicant. The path analysis revealed that 1000 seed weight had highest positive direct relationship with seed yield (1.057), followed by total chlorophyll (0.559) and branches/plant (0.520). Traits showing high negative direct effect on seed yield were leaf carotenoid (0.749), seed size (0.678) and days to owering (0.377). Total chlorophyll exerted strongest direct positive effect (0.722) on harvest index, followed by seed yield (0.505) and seed protein (0.245). # 2006 Elsevier B.V. All rights reserved.
Keywords: Chenopodium quinoa; Seed yield; Harvest index; Protein; Carotenoid; Heritability; Genetic advance; Correlation; Selection approaches

1. Introduction Quinoa (Chenopodium quinoa Willd.), an Andean crop, has recently gained worldwide attention because of its ability to grow in various stress conditions like soil salinity, acidity, drought, frost, etc. (Vacher, 1998; Jensen et al., 2000; Bhargava et al., 2003a, 2006a; Jacobsen et al., 2003). Apart from this, its grain is a rich source of a wide range of minerals, vitamins, oil containing large amounts of linoleate, linolenate and natural antioxidants (Koziol, 1992; Repo-Carrasco et al., 2003) and high quality protein containing ample amounts of sulphur rich amino acids (Koziol, 1992). These benets coupled with increasing demand for quinoa and insufcient supply from quinoa producing countries of South America necessitates introduction of the crop to newer areas. Quinoa was introduced in England in 1970 and later in Denmark. The crop has gained rm ground in the region and many varieties suited to local conditions have been released (Mastebroek and Limburg, 1996; Jacobsen, 2003). In the United

* Corresponding author. Tel.: +91 522 2205831; fax: +91 522 2205836. E-mail address: atul_238@rediffmail.com (A. Bhargava). 0378-4290/$ see front matter # 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.fcr.2006.10.001

States, work with quinoa started in the early 1980s and presently its consumption is approximately 1500 t/ha. However, in Asia, concerted research efforts on quinoa are lacking and the crop has not been provided due importance. The Indo-Gangetic plain (IGP), a region of land covering large areas of India, Pakistan, Nepal and Bangladesh, is characterized by fertile soils, favourable climate and an abundant supply of water (Aggarwal et al., 2004). In India, the states of Uttar Pradesh and Bihar constitute an important transect of the IGP which is characterized by hot, subhumid climate with bulk of rainfall received during the monsoon season. The winters in the transect are cool and nearly uniform climatic features are present in these states with slight deviations. However, this area is also amongst the most prone areas of the world to degradation of natural resources due to intense human activity. There is growing concern about the decline in soil fertility, changes in water table depth, deterioration in the quality of irrigation water, and rising salinity in the region (Joshi and Tyagi, 1994; Aggarwal et al., 2004). A large portion of the population in this region has little access to protein rich diet since wheat and rice are the principal food crops grown and consumed in the area. The increasing

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population demands an increase in food production along with a shift towards environmentally sound sustainable agriculture. There is a need for cultivation of crops that requires minimum inputs, but can counter the nutritional deciency prevalent in the general population of this region. Quinoa can be termed underutilized, especially for India, since inspite of its wide adaptability, rusticity and nutritional superiority, its commercial potential has remained untapped. Quinoas highly proteinaceous grain can help to make diets more balanced in this region and can play an important role in combating silent hunger among poor populations in India who have little access to protein rich diet. Quinoas ability to produce high protein grains under ecologically extreme conditions makes it important for the diversication of future agricultural systems, especially in high altitude area of the Himalayas and North Indian Plains. The worldwide popularity of quinoa and initial promising reports from India (Bhargava et al., 2003a) makes it imperative for its evaluation in this transect of the IGP to explore its potential as an alternative winter crop in the region. The main aim of quinoa breeders is the development of cultivars with high grain yield and quality components, adapted to diverse agro-climatic regions. Inspite of the immense nutritive importance of the crop, not much work has been done for its genetic improvement leading to lack of information on many aspects. Breeding a crop for new and targeted environments requires the use of a range of cultivars/genotypes since it allows us to quantify intraspecic variability for different traits and their interactions. Genetic variability in the base population plays a very important role in any cropbreeding program. The extent of diversity present in the germplasm determines the limits of selection for improvement. The characters of economic importance are generally quantitative in nature and exhibit considerable degree of interaction with the environment. Thus, it becomes imperative to compute the variability present in the material and its partitioning into genotypic, phenotypic and environmental ones. Improvement of yield requires an in-depth knowledge of the magnitude of variation present in the available germplasm, interdependence of quantitative characters with yield, extent of environmental inuence on these factors, heritability and genetic gain of the material. Correlation coefcients show relationships among various traits along with the degree of linear relation between these characters. But, correlation studies provide incomplete information on the relative importance of the direct and indirect effects of individual traits on yield. Thus, simple correlations are less likely to provide a clear picture of the importance of each component trait in determining yield. In such condition, it becomes necessary to study the path coefcient analysis, which takes into account not only the causal relationship, but also the degree of relationship among the traits. Path analysis, also known as standardized partial-regression coefcient, partitions the correlation coefcients into direct and indirect effects and thereafter allows the separation of direct inuence of each trait on yield from the indirect effects caused by mutual association among the traits themselves (Garcia del Moral et al., 2003). Reports on variability and association among different traits in

quinoa are rare, based on few yield components and are based on experiments carried out in America and Europe (Risi and Galwey, 1989; Rojas et al., 2003). Detailed experimental results on morphological and qualitative variability, and correlation and path analysis from Asia, particularly the Indo-Gangetic plains are altogether absent. Therefore, the present study was conducted with the following objectives: (a) To ascertain the suitability of quinoa as a new winter crop in the Indo-Gangetic plains of northern India. (b) To evaluate the morphological and qualitative characteristics of quinoa germplasm. (c) To elucidate interrelationships among yield and yield components using correlation analysis and supplementing correlation results using path coefcient analysis. (d) To determine the selection criteria for increasing seed yield and harvest index. 2. Materials and methods 2.1. Experimental site The experiment was conducted at the experimental eld of National Botanical Research Institute, Lucknow. The experimental site is situated at an altitude of 120 m above sea level at 26.5 8N latitude and 80.5 8E longitude. In the Indo-Gangetic plains of North India there are two main crop seasons, summer (kharifMarch to July) and winter (rabiOctober to February). Chenopodium grows mostly during the rabi season. During the rabi season, the minimum and maximum temperature ranges from 2.5 to 19 and 14 to 29 8C, respectively. 2.2. Experimental set up Field experiments were conducted in the crop years 2002 2003 and 20032004 on sandy loam soil at the experimental site. The weather parameters prevailing during both the experimental years have been provided in Table 1. Twentyseven germplasm lines of C. quinoa and two lines of its closest relative, C. berlandieri subsp. nuttalliae (all introduced) were sown as a rabi crop in mid November in both the years and evaluated for various morphological and quality traits. The colour of the seeds was determined as translucent, white, yellow, orange, red, ochre, brown purple or black. The rst four were categorized as light while the latter ve as dark (Table 2). All the lines selected were tetraploid (2n = 36), and had been collected from varying altitudes and locations (Table 2). The eld was disc ploughed and then harrowed and raked to obtain a good seed bed before sowing. The experimental design was a randomized block with three replications. The plot size for each replication was 2 m 2 m. Each plot had 6 rows spaced 30 cm apart and each row had 10 plants separated at 20 cm from each other. For the whole crop season, weeding followed by hoeing was done at an interval of 15 days. Irrigation was provided as and when needed. No chemical fertilizer was applied either before or during the experiment. These was primarily done to ascertain the potential of the crop for subsistence agriculture

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Table 1 Weather conditions during the rst and second experiments Temperature (8C) Maximum Experiment I (20022003) November 24 December 18 January 15 February 24 March 30 April 38 Experiment II (20032004) November 28 December 21 January 10 February 25 March 34 April 37 Minimum 16 11 7 11 16 23 14 11 8 12 17 23 Mean 20 15 11 18 23 30 21 16 9 19 26 30 14 10 8 12 13 15 12 11 7 11 13 16 3 4 3 6 7 9 2 4 3 4 5 7 Dew point (8C) Wind (km/h)

quinoa and the plant survives mainly on residues of manure or fertilizer from the previous crop (Aguilar and Jacobsen, 2003). Each germplasm line was sown in a separate plot and thinning was done to maintain plant density within rows. Irrigation was applied as and when needed. No fungicide or insecticide was used during the experiment. Hand weeding was carried out once in 20 days to remove unwanted plants. 2.3. Parameters estimated Ten plants from the central rows of each germplasm line in each replication were randomly tagged and data were recorded on these plants for the following morphological traits: (a) Days to owering: the number of days from the date of emergence to the date at which about 50% of the plants in a plot showed blooming. (b) Days to maturity: days to maturity was taken from date of emergence to the date when the crop was ready for harvesting, i.e. seeds had become mature and the plant had started drying. (c) Plant height (cm): the average height from the ground level to the tip of the inorescence on the main stem at the time of harvesting was measured.

since a large chunk of the farmers in the region are nancially weak and seek crops with fewer inputs. We tried to emulate the same type of cultivation practice as followed in the Peruvian altiplano, where farmers do not use any kind of fertilizers for
Table 2 Germplasm lines, their source, origin and seed colour Germplasm line C. quinoa Willd. CHEN 58/77 C. quinoa Willd. CHEN 67/78 C. quinoa Willd. CHEN 71/78 C. quinoa Willd. CHEN 33/84 C. quinoa Willd. CHEN 84/79 C. quinoa Willd. CHEN 92/91 C. quinoa Willd. CHEN 7/81 C. quinoa Willd. PI 614938 C. quinoa Willd. PI 478408 C. quinoa Willd. PI 478414 C. quinoa Willd. PI 596498 C. quinoa Willd. Ames 13219 C. quinoa Willd. Ames 13719 C. quinoa Willd. PI 587173 C. quinoa Willd. PI 510532 C. quinoa Willd. PI 614883 C. quinoa Willd. PI 584524 C. quinoa Willd. Ames 22156 C. quinoa Willd. Ames 13762 C. quinoa Willd. PI 614881 C. quinoa Willd. PI 510537 C. quinoa Willd. PI 510547 C. quinoa Willd. Ames 22158 C. quinoa Willd. PI 510536 C. quinoa Willd. PI 478410 C. quinoa Willd. PI 433232 C. quinoa Willd. Ames 21909 C. berlandieri subsp. nuttalliae PI 568155 (Saff.) Wilson and Heiser C. berlandieri subsp. nuttalliae PI 568156 (Saff.) Wilson and Heiser
a b

Source IPK Gatersleben, IPK Gatersleben, IPK Gatersleben, IPK Gatersleben, IPK Gatersleben, IPK Gatersleben, IPK Gatersleben, USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA USDA Germany Germany Germany Germany Germany Germany Germany

Statusa Cultivar Cultivar Landrace Cultivated Cultivated Cultivated Cultivar Cultivated Cultivated Landrace Cultivated Cultivar Landrace Landrace Landrace

Origina Puno, Peru Bolivia Cuzco, Peru Columbia Oruro, Bolivia La Paz, Bolivia La Paz, Bolivia Cuzco, Peru La Paz, Bolivia New Mexico, USAb Jujuy, Argentina Peru Jujuy, Argentina Chile Chile New Mexico, USAb Jujuy, Argentina Peru Peru Chile Peru La Paz, Bolivia Chile Oruro, Bolivia Mexico Mexico

Altitudea (m) 4000 3200 3800 3800 3030 3700 3000 3800 3870 1680 2700

Seed colour Light Dark Light Light Light Light Light Light Light Dark Light Light Light Light Light Light Light Light Light Light Dark Dark Light Dark Light Light Light Dark Dark

From germplasm database record of the collection or donation site, which might not indicate biological adaptation. Since C. quinoa Willd. is not native to USA, this indicates the location of a germplasm donation, and might not indicate biological adaptation.

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(d) Leaf area (cm2): leaf area was measured for three positions, viz. top, middle and lower, using the leaf area meter of Delta T Devices Ltd., when the plant was in full bloom. (e) Primary branches/plant: the total number of branches growing from the main stem at different node positions, including the basal branches. (f) Inorescence length (cm): the mean length of three inorescence was taken randomly from different positions. (g) Inorescence/plant: the number of inorescence per plant was counted at the time of harvest. (h) Seed size (mm): the seed size was measured following the method suggested by Bertero et al. (2004). (i) Thousand seed weight (g): a sample of 1000 seeds from the bulked seed of each line was weighted. (j) Dry weight/plant (g): whole plants excluding the roots, secondary branches and leaves were sun dried and weighted. (k) Harvest index: this was calculated by the following formula: seed yield=plant dry weight=plant (l) Seed yield (t/ha): the seed of all the plants of each plot were bulked and weighed and the seed yield/plot was then converted to tonnes per hectare (t/ha). harvest index Apart from this, four quality traits were also estimated namely: (a) Total chlorophyll (mg/g): total chlorophyll was calculated from fresh leaves collected from the tagged plants at the 9week-old stage as per the method proposed by Jensen (1978). (b) Leaf carotenoid (mg/kg): the carotenoid content of leaves was determined from fresh leaves collected from the tagged plants at the 9-week-old stage as per the method described by Jensen (1978). (c) Seed carotenoid (mg/kg): seed carotenoid content was estimated from the bulked seed of each line according to Jensen (1978). (d) Seed protein (%): the protein content was estimated from the bulked seed of each germplasm line following the method suggested by Peterson (1977). 2.4. Statistical analysis The raw data was compiled by taking the means of all the plants taken for each treatment and replication for different traits in both the experimental years. The pooled means of both the years were subjected to further statistical and biometrical analysis. Simple statistical parameters, viz. mean, standard error, variance and coefcient of variation were analyzed according to Singh and Chaudhary (1985). Heritability is dened the fraction of genetic variation in relation to environmental variance. For computing broad sense heritability, the following formula proposed by Singh and Chaudhary (1985) was used: H s2g s2 p

where s2g = genotypic variance and s2p = phenotypic variance. Genetic advance is referred as the difference between the genotypic mean of selected lines and genotypic mean of population. Genetic advance as percentage of mean was calculated by the following formula (Singh and Chaudhary, 1985): genetic advance % genetic advance trait 100 mean trait

Correlation analysis was performed to determine the relationships between yield and all the component traits, both at genotypic and phenotypic levels according to Johnson et al. (1955a). The phenotypic level included both genotypic and environmental factors, while genotypic analysis focused strictly on genetic effects and excluded the environmental ones. Path coefcients were then computed (Dewey and Lu, 1959) to separate the direct and indirect effects of correlation coefcient. A signicant positive correlation between a trait and yield, high positive direct effect by that trait on yield and minimal negative indirect effect by that trait on yield via other traits, were the three factors that categorized the trait as effective for yield improvement. 3. Results 3.1. Variability studies The analysis of variance for two separate years revealed signicant differences among the strains for all the 16 characters, which validated further statistical and genetic analysis (Table 3). These results indicate the presence of a high degree of morphological and qualitative variation among the lines studied. The year variety/line interaction was non-signicant for all the traits except for inorescence length (Table 3).
Table 3 Mean squares of the analysis of variance for 12 morphological and 4 quality traits in Chenopodium Traits Mean sum of squares Year I Days to owering Days to maturity Plant height (cm) Leaf area (cm2) Branches/plant Inorescence length (cm) Inorescence/plant Seed size (mm) 1000 seed weight (g) Dry weight/plant (g) Harvest index Seed yield (t/ha) Total chlorophyll (mg/g) Leaf carotenoid (mg/kg) Seed carotenoid (mg/kg) Seed protein (%)
*

Year II
**

G year
**

114.26 513.15** 4003.66** 169.25** 96.12** 5.46** 5196.60** 0.14** 2.19** 444.29** 0.32** 23.13** 0.337** 28559.40** 2.49** 20.16**

131.55 569.12** 3840.29** 188.11** 105.54** 5.02 5346.18** 0.15** 2.30** 418.82** 0.35** 26.29** 0.349** 29014.54** 2.31** 19.44**

149.20 484.62 1019.06 119.24 101.50 7.19* 4623.10 0.12 2.04 384.10 0.19 19.79 0.302 20126.43 1.93 13.32

P = 0.05;

**

P = 0.01.

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Table 4 Mean performance of 29 lines for 12 morphological traits in Chenopodium

Germplasm lines Origin Days to owering Days to maturity 117.67 119.44 131.67 144.00 121.67 123.22 133.78 109.33 109.33 134.11 129.00 129.98 120.28 125.78 157.11 109.89 127.00 126.00 132.44 127.22 124.00 131.78 131.11 115.22 126.78 130.00 152.44 163.33 152.33 Plant height (cm) 45.41 59.63 46.33 42.33 86.97 77.49 123.56 11.27 17.67 78.98 65.87 53.96 115.52 101.03 144.03 54.89 115.89 106.44 123.72 113.00 100.00 66.67 80.27 31.05 101.10 108.66 82.44 139.44 135.44 Leaf area (cm2) 15.71 6.12 26.94 9.46 17.47 24.69 22.14 5.67 8.93 21.53 20.82 11.75 25.03 30.91 22.02 12.33 29.64 26.16 5.00 25.00 14.39 16.02 23.25 4.42 17.29 23.01 25.87 21.44 13.53 Primary branches/ plant 16.56 16.70 15.44 16.96 22.11 14.06 28.00 10.00 8.55 20.55 17.33 19.21 27.74 16.74 25.55 21.89 25.00 20.44 23.00 24.56 25.44 14.11 21.24 17.53 22.61 20.89 21.00 35.74 29.11 Inorescence Inorescence/ Seed length (cm) plant size (mm) 2.93 1.71 3.39 2.42 1.00 2.25 4.09 1.07 0.84 1.60 2.47 2.64 2.67 2.25 2.24 3.61 2.51 1.60 4.31 3.01 1.44 2.08 3.85 1.79 0.90 4.54 2.12 6.47 4.77 41.19 91.63 127.73 13.85 117.78 64.11 141.55 11.67 14.65 106.48 90.33 114.66 98.00 68.50 138.22 45.89 137.55 85.55 136.44 114.22 136.00 68.92 40.29 21.03 118.33 74.22 132.22 114.78 103.39 1.58 1.34 1.97 1.57 2.21 2.01 2.09 1.73 2.17 1.81 2.03 2.06 2.15 2.01 1.51 1.73 1.58 1.93 1.83 2.05 1.78 1.82 1.95 1.93 1.80 1.77 1.83 1.58 1.65 1000 seed weight (g) 1.81 0.78 2.85 2.07 3.57 3.70 3.65 1.87 2.87 3.03 3.08 3.54 3.65 4.09 1.25 1.77 3.02 3.51 2.75 2.94 2.71 3.13 3.17 2.34 2.63 2.28 3.31 1.28 1.37 Dry weight/ plant (g) 6.31 5.75 7.21 3.84 10.47 10.21 28.00 1.11 1.26 14.00 19.89 15.08 32.03 15.47 52.89 3.03 29.86 17.21 35.21 24.16 13.02 12.67 12.70 1.38 29.00 13.11 15.97 28.94 15.05 Harvest index 1.07 0.74 1.43 1.40 1.32 0.88 1.41 1.06 1.19 1.25 0.79 0.73 0.99 0.81 0.29 0.97 0.90 1.21 0.94 1.34 1.32 1.33 1.18 1.28 0.43 1.09 1.15 0.26 0.65 Seed yield (t/ha) 2.11 3.75 3.27 1.33 3.44 2.25 9.83 0.32

C. C. C. C. C. C. C. C.

quinoa quinoa quinoa quinoa quinoa quinoa quinoa quinoa

CHEN 58/77 CHEN 67/78 CHEN 71/78 CHEN 33/84 CHEN 84/79 CHEN 92/91 CHEN 7/81 PI 614938

C. quinoa PI 478408 C. quinoa PI 478414 C. quinoa PI 596498 C. quinoa Ames 13219 C. quinoa Ames 13719 C. quinoa PI 587173 C. quinoa PI 510532 C. quinoa PI 614883 C. quinoa PI 584524 C. quinoa Ames 22156 C. quinoa Ames 13762 C. quinoa PI 614881 C. C. C. C. C. C. C. C.

quinoa PI 510537 quinoa PI 510547 quinoa Ames 22158 quinoa PI 510536 quinoa PI 478410 quinoa PI 433232 quinoa Ames 21909 berlandieri subsp. nuttalliae PI 568155 C. berlandieri subsp. Mexico nuttalliae PI 568156

73.55 Puno, Peru 74.55 Bolivia 79.33 101.55 Cuzco, Peru 86.00 Columbia 81.89 85.11 Oruro, 71.00 Bolivia La Paz, 71.33 Bolivia La Paz, 83.66 Bolivia Cuzco, Peru 83.77 La Paz, 81.99 Bolivia New Mexico, 82.21 USA Jujuy, 85.33 Argentina Peru 86.67 Jujuy, 70.78 Argentina Chile 81.33 Chile 80.55 New Mexico, 79.33 USA Jujuy, 87.11 Argentina Peru 84.33 Peru 82.11 Chile 80.89 Peru 73.78 La Paz, Bolivia 82.77 Chile 81.00 Oruro, Bolivia 82.55 Mexico 91.33 85.33

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0.47 6.07 3.93 2.80 9.33 3.17 1.68 1.00 6.60 5.03 8.50 8.25 4.39 4.70 4.85 0.67 3.13 3.56 9.08 2.01 2.32

Mean S.E. CD (5%) CD (1%) CV

81.76 1.18 129.51 2.51 2.41 5.14 3.26 6.93 7.82 10.44

83.76 6.79 18.15 1.44 20.62 1.08 2.64 0.24 13.90 2.94 2.21 0.49 18.76 3.97 2.98 0.66 43.67 42.75 28.32 49.62

88.59 7.81 1.84 0.03 2.69 0.15 16.37 2.24 1.01 0.06 4.06 0.52 15.99 0.06 0.30 4.58 0.12 1.06 21.57 0.08 0.41 6.18 0.17 1.43 47.48 11.41 31.97 73.85 32.16 68.34

A. Bhargava et al. / Field Crops Research 101 (2007) 104116 Table 5 Mean performance of 29 lines for 4 quality traits in Chenopodium Germplasm lines C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. C. quinoa CHEN 58/77 quinoa CHEN 67/78 quinoa CHEN 71/78 quinoa CHEN 33/84 quinoa CHEN 84/79 quinoa CHEN 92/91 quinoa CHEN 7/81 quinoa PI 614938 quinoa PI 478408 quinoa PI 478414 quinoa PI 596498 quinoa Ames 13219 quinoa Ames 13719 quinoa PI 587173 quinoa PI 510532 quinoa PI 614883 quinoa PI 584524 quinoa Ames 22156 quinoa Ames 13762 quinoa PI 614881 quinoa PI 510537 quinoa PI 510547 quinoa Ames 22158 quinoa PI 510536 quinoa PI 478410 quinoa PI 433232 quinoa Ames 21909 berlandieri subsp. nuttalliae PI 568155 C. berlandieri subsp. nuttalliae PI 568156 Mean S.E. CD (5%) CD (1%) CV Origin Puno, Peru Bolivia Cuzco, Peru Columbia Oruro, Bolivia La Paz, Bolivia La Paz, Bolivia Cuzco, Peru La Paz, Bolivia New Mexico, USA Jujuy, Argentina Peru Jujuy, Argentina Chile Chile New Mexico, USA Jujuy, Argentina Peru Peru Chile Peru La Paz, Bolivia Chile Oruro, Bolivia Mexico Mexico Total chlorophyll (mg/g) 1.03 1.70 1.82 0.55 1.12 1.68 1.92 1.16 1.19 1.86 1.65 1.32 1.36 1.85 1.34 1.25 2.04 1.86 1.60 1.42 1.59 1.22 1.06 1.09 1.43 1.51 1.55 1.17 1.20 1.43 0.06 0.12 0.16 23.07 Leaf carotenoid (mg/kg) 389.83 531.03 534.80 230.23 414.73 521.83 632.40 338.23 330.03 588.23 551.07 421.03 466.13 580.43 483.13 434.67 669.56 611.83 519.90 481.23 511.77 416.30 414.63 371.80 480.07 479.47 504.07 601.90 528.50 484.09 18.37 37.62 50.75 20.42 Seed carotenoid (mg/kg) 1.73 3.12 3.15 1.69 2.30 2.00 3.30 2.84 2.74 3.88 2.68 2.02 1.75 3.86 2.06 3.15 2.87 2.81 2.08 3.33 3.82 2.35 2.40 2.84 1.97 2.13 3.15 5.52 4.73 2.83 0.16 0.32 0.44 31.80

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Seed protein (%) 13.22 21.02 19.37 16.92 18.84 13.93 17.31 17.83 15.23 17.86 15.09 12.55 17.71 14.66 14.51 19.48 13.01 14.24 15.47 13.89 19.78 20.43 16.09 20.39 13.08 14.23 16.20 13.28 14.82 16.22 0.47 0.96 1.29 15.90

The mean values for different morphological and quality traits are presented in Tables 4 and 5. Out of 29 lines, only 11 lines showed above average seed yield. Seed yield ranged from 0.32 to 9.83 t/ha with C. quinoa PI 614938 showing the lowest yield. Highest seed yield was shown by C. quinoa CHEN 7/81 (9.83 t/ha), followed by C. quinoa Ames 13719 (9.33 t/ha) and C. quinoa Ames 21909 (9.08 t/ha). These lines also showed above average mean performance for most of the morphological traits. The three lowest yielding lines, viz. C. quinoa PI 614938, C. quinoa PI 478408 and C. quinoa PI 510536 showed low values for most morphological traits, total chlorophyll and leaf carotenoid. Five germplasm lines of quinoa (CHEN 58/77, Ames 13219, PI 510532, PI 478410 and PI 433232) were poor both in terms of yield and quality. Both the lines of C. berlandieri subsp. nuttalliae (PI 568155 and PI 568156) exhibited high values for most of the morphological traits but were low yielding (2.01 and 2.32 t/ha, respectively) (Table 4). Harvest index ranged from 0.26 to 1.43, with C. quinoa CHEN 71/78 showing the highest value (1.43), followed by C. quinoa CHEN 7/81 (1.41) and C. quinoa CHEN 33/84 (1.40). Seed protein among the lines ranged from 12.55 to 21.02% with an average of 16.22 0.47%, while seed carotenoid was in the

range of 1.695.52 mg/kg with a mean of 2.83 0.16 mg/kg (Table 5). Highest seed protein was found in C. quinoa CHEN 67/78 (21.02%), followed by C. quinoa PI 510547 (20.43%) and C. quinoa PI 510536 (20.39%) all of which had dark coloured seeds. The carotenoid content in the leaves ranged from 230.23 mg/kg for C. quinoa CHEN 33/84 to 669.56 mg/ kg for C. quinoa PI 584524, and was comparatively higher than that found in the seeds. Seventy percent of the lines having high leaf carotenoid also had high seed carotenoids. Among the 12 morphological traits, dry weight/plant, seed yield and inorescence length exhibited high values for coefcient of variability while for quality traits these values were relatively low, the highest being for seed carotenoid (31.80%) and the lowest for seed protein (15.90%) (Table 5). The values for variance, coefcient of variation, heritability and genetic gain for different morphological and quality traits are presented in Table 6. Phenotypic variance denoting total variance was maximum for inorescence/plant among morphological traits and for leaf carotenoid among the quality traits. The phenotypic coefcient of variation (PCV) values for all the traits were higher than the corresponding genotypic values (GCV) values though had small differences. Among all

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Table 6 Range, variance, coefcient of variation, heritability and genetic gain for various morphological and quality traits in Chenopodium Traits Days to owering Days to maturity Plant height (cm) Leaf area (cm2) Primary branches/plant Inorescence length (cm) Inorescence/plant Seed size (mm) 1000 seed weight (g) Dry weight/plant (g) Harvest index Seed yield (t/ha) Total chlorophyll (mg/g) Leaf carotenoid (mg/kg) Seed carotenoid (mg/kg) Seed protein (%) Range 70.78101.55 109.33163.33 11.27144.03 4.4230.91 8.5535.74 0.846.47 11.67141.55 1.342.21 0.784.09 1.1152.89 0.261.43 0.329.83 0.552.04 230.23669.56 1.695.52 12.5521.02 s2p 42.36 185.75 1347.37 62.47 35.20 1.750 1824.32 0.053 0.759 150.63 0.106 7.71 0.122 10452.16 0.833 6.73 s2g 40.43 182.29 1333.55 59.16 33.59 1.73 1744.54 0.045 0.732 144.03 0.105 7.68 0.109 9441.26 0.813 6.68 s2e 1.93 3.46 13.82 3.31 1.61 0.018 79.78 0.008 0.027 6.60 0.001 0.032 0.013 1010.90 0.020 0.052 PCV 7.96 10.53 43.82 43.53 28.76 50.12 48.20 12.53 32.39 74.95 32.15 68.33 24.48 21.11 32.18 15.98 GCV 7.77 10.43 43.59 42.36 28.10 49.85 47.14 11.49 31.80 73.29 32.05 68.19 23.15 20.07 31.78 15.92 Heritability (%) 95.45 98.14 98.97 94.69 95.42 98.92 95.63 84.90 96.44 95.62 99.42 99.59 89.34 90.33 97.60 99.23 Genetic gain (%) 15.65 21.27 89.35 84.94 56.56 102.08 94.98 21.88 64.34 147.68 65.84 140.19 44.95 39.29 64.84 32.69

s2p: phenotypic variance; s2g: genotypic variance; s2e: environmental variance; PCV: phenotypic coefcient of variation; GCV: genotypic coefcient of variation.

the 16 traits studied, dry weight/plant, seed yield and inorescence length recorded high coefcient of variation values. Broad sense heritability was high and exceeded 80% for all the traits. Genetic gain as percent of mean was highest for seed yield (140.19%), followed by dry weight/plant (133.62%) and inorescence length (102.08%). Seed protein showed lowest genetic gain among quality traits (32.69%) while for morphological traits the lowest value was for days to owering (15.65%). 3.2. Correlation studies The genotypic and phenotypic correlation coefcients between various traits are presented in Table 7. The genotypic correlation values were slightly higher than their corresponding phenotypic values, which might be due to the modied effect of environment on character association at the genetic level. All morphological traits except days to owering, days to maturity and inorescence length exhibited signicant positive association with seed yield, the maximum value was recorded for inorescence/plant. Among the quality traits, total chlorophyll showed maximum correlation with seed yield both at phenotypic and genotypic levels (0.499** and 0.509**, respectively). Traits like days to owering, days to maturity, plant height, leaf area, primary branches/plant, inorescence/ plant and dry weight/plant showed positive association amongst themselves, which were highly signicant. The association of leaf area with all the traits except harvest index and seed protein was positive. Seed size was negatively correlated with all the morphological traits except for leaf area, 1000 seed weight, harvest index and seed yield. The association of leaf carotenoid with total chlorophyll and seed carotenoid was positive and highly signicant. Leaf carotenoid also showed positive correlation with all morphological traits except seed size. An interesting observation relates to the negative association exhibited by seed protein with 13 traits. Of these the relationship with days to maturity, plant height, leaf area, inorescence length and dry weight/plant were signicant,

while harvest index was the only trait exhibiting signicant positive association with seed protein. Both seed carotenoid and seed protein were negatively correlated with seed yield, though the values were non-signicant (Table 7). 3.3. Path analysis In the present study, path coefcient analysis has been conducted taking harvest index and seed yield as dependent variables. The direct and indirect effects of various traits on seed yield are provided in Table 8. The path analysis revealed that 1000 seed weight had highest positive direct relationship with seed yield (1.057), followed by total chlorophyll (0.559) and branches/plant (0.520). Some traits showing high negative direct effect were leaf carotenoid (0.749), seed size (0.678) and days to owering (0.377). Days to owering exerted negative indirect inuence on seed yield via all the morphological traits except seed size and harvest index. Likewise, leaf area also negatively indirectly inuenced seed yield through most of the morphological traits except plant height and harvest index. Among the quality traits, seed protein showed negative indirect effect on seed yield through all the traits barring harvest index and seed carotenoid, though the values of indirect effects were small. The direct and indirect effects of various characters on harvest index are given in Table 9. Total chlorophyll exerted strongest direct positive effect (0.722) on harvest index, followed by seed yield (0.505) and seed protein (0.245). Seed yield also indirectly and positively inuenced harvest index through all the traits except leaf and seed carotenoid. Highest negative direct was exhibited by leaf carotenoid (1.118), dry weight/plant (0.638) and days to maturity (0.058). Seed protein and dry weight/plant negatively indirectly inuenced harvest index through all the morphological traits while inorescence/plant and leaf carotenoid were the other traits exercising negative inuence on harvest index through majority of other traits. Plant height and leaf area showed positive direct

Table 7 Genotypic and phenotypic correlation coefcients among 16 traits in Chenopodium Traits Days to Days to Plant owering maturity height (cm) G 0.201 P 0.199 G P G P G P G P G P 0.178 0.177 Leaf area (cm2) Branches/ Inorescence Inorescence/ Seed plant length (cm) plant size (mm) 0.154 0.153 0.246 0.244 0.522** 0.520** 0.484** 0.483** 0.169 0.166 0.606** 0.600** 0.613** 0.607** 0.331** 0.325* 0.481** 0.478** 0.719** 0.714** 0.422** 0.414** 0.644** 0.640** 0.203 0.205 0.277* 0.267* 0.062 0.056 1000 seed Dry weight Harvest weight (gm) /plant (g) index 0.491** 0.485** 0.119 0.114 0.486** 0.480** 0.389** 0.384** 0.542** 0.536** 0.825** 0.821** 0.402** 0.395** 0.663** 0.655** 0.272* 0.273* 0.706** 0.699** 0.086 0.079 0.037 0.041 0.267* 0.267* 0.070 0.069 Total Leaf Seed Seed chlorophyll carotenoid caro-enoid protein (mg/g) (mg/kg) (mg/kg) (%) 0.509** 0.499** 0.146 0.138 0.506** 0.007 0.498** 0.006 0.100 0.103 0.309* 0.300* 0.669** 0.657** 0.618** 0.604** 0.515** 0.507** 0.304* 0.299* 0.718** 0.703** 0.044 0.039 0.205 0.199 0.129 0.128 0.018 0.017 0.237 0.236

Seed yield/plant (g)

0.507** 0.433** 0.417** 0.504** 0.428** 0.413**

Days to owering

0.712** 0.492** 0.339** 0.474** 0.702** 0.488** 0.332* 0.467** 0.611** 0.310* 0.608** 0.305* 0.596** 0.590**

Days to maturity

0.356** 0.226 0.343* 0.224 0.104 0.102 0.254* 0.241 0.165 0.161 0.199 0.190 0.003 0.002 0.035 0.036 0.504** 0.498** 0.143 0.143 0.264* 0.261* 0.158 0.155 0.849** 0.817**

0.381** 0.042 0.378** 0.043 0.410** 0.409** 0.022 0.022 0.367** 0.362** 0.248 0.247 0.237 0.234 0.423** 0.408** 0.426** 0.420** 0.516** 0.513** 0.356** 0.348** 0.533** 0.526** 0.096 0.101 0.047 0.046 0.591** 0.580** 0.131 0.127 0.396** 0.386** 0.299* 0.302* 0.018 0.017

0.369** 0.296* 0.367** 0.293* 0.298* 0.295* 0.142 0.137 0.382** 0.380** 0.379** 0.375** A. Bhargava et al. / Field Crops Research 101 (2007) 104116

Plant height (cm)

0.503** 0.849** 0.498** 0.843** 0.304* 0.301*

Leaf area (cm2)

Branches/plant

0.442** 0.223 0.435** 0.221 0.388** 0.266* 0.386** 0.265* 0.242 0.238 0.169 0.164 0.231 0.229 0.159 0.157 0.012 0.011 0.115 0.114 0.433** 0.429** 0.500** 0.498** 0.076 0.075

Inorescence length (cm) G P Inorescence/plant G P G P G P G P G P G P G P G P

**

Seed size G (mm)

1000 seed weight (g)

Dry weight/plant (g)

0.501** 0.015 0.498** 0.017 0.271* 0.264* 0.865** 0.862** 0.171 0.169 0.242 0.236

Harvest index P

Total chlorophyll (mg/g)

Leaf carotenoid (mg/kg)

0.508** 0.228 0.496** 0.225 0.066 0.067 111 P = 0.01.

Seed carotenoid (mg/kg)

G: genotypic correlation coefcient; P: phenotypic correlation coefcient. *P = 0.05,

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Table 8 Path coefcient analysis showing direct (in bold) and indirect effects of 15 traits over seed yield in Chenopodium Traits Days to Days to Plant owering maturity height (cm) 0.377 0.254 0.176 0.121 0.169 0.088 0.128 0.022 0.043 0.139 0.025 0.052 0.036 0.046 0.085 0.089 0.125 0.076 0.039 0.074 0.065 0.060 0.044 0.028 0.068 0.047 0.005 0.039 0.046 0.037 0.020 0.009 0.031 0.001 0.008 0.032 0.010 0.023 0.006 0.019 0.040 0.001 0.022 0.007 0.010 Leaf area (cm2) 0.037 0.034 0.086 0.109 0.033 0.050 0.046 0.028 0.055 0.044 0.002 0.058 0.068 0.016 0.041 Branches/ Inorescence Inorescence/ Seed plant length (cm) plant size (mm) 0.242 0.305 0.434 0.157 0.520 0.310 0.330 0.084 0.073 0.320 0.188 0.048 0.263 0.219 0.114 0.049 0.104 0.097 0.034 0.121 0.200 0.041 0.040 0.053 0.054 0.049 0.009 0.061 0.077 0.053 0.042 0.061 0.091 0.053 0.082 0.026 0.127 0.001 0.020 0.089 0.030 0.075 0.091 0.031 0.020 0.042 0.241 0.071 0.172 0.111 0.135 0.002 0.678 0.570 0.058 0.286 0.089 0.030 0.115 0.008 Genotypic 1000 seed Dry weight/ Harvest Total Leaf Seed Seed weight (g) plant (g) index chlorophyll carotenoid carotenoid protein Correlation (mg/kg) (mg/kg) (%) (mg/g) 0.126 0.248 0.037 0.533 0.151 0.279 0.167 0.875 1.057 0.039 0.450 0.419 0.216 0.244 0.121 0.187 0.260 0.396 0.193 0.319 0.131 0.339 0.041 0.018 0.481 0.248 0.144 0.241 0.007 0.208 0.031 0.170 0.183 0.010 0.164 0.111 0.106 0.189 0.191 0.231 0.408 0.008 0.121 0.076 0.214 0.082 0.023 0.198 0.298 0.054 0.026 0.330 0.073 0.221 0.167 0.010 0.559 0.461 0.135 0.043 0.075 0.232 0.501 0.463 0.385 0.228 0.538 0.033 0.153 0.375 0.203 0.647 0.749 0.380 0.170 0.017 0.048 0.038 0.018 0.057 0.050 0.031 0.022 0.030 0.002 0.022 0.031 0.066 0.129 0.009 0.010 0.201 0.012 0.178 0.016 0.507** 0.016 0.433** 0.009 0.417** 0.011 0.155 0.006 0.613** 0.001 0.277* 0.005 0.491** 0.018 0.486** 0.021 0.267** 0.003 0.509** 0.009 0.506** 0.003 0.007 0.042 0.018

Days to owering Days to maturity Plant height (cm) Leaf area (cm2) Branches/plant Inorescence length (cm) Inorescence/plant Seed size (mm) 1000 seed weight (g) Dry weight/plant (g) Harvest index Total chlorophyll (mg/g) Leaf carotenoid (mg/kg) Seed carotenoid (mg/kg) Seed protein (%)
*

A. Bhargava et al. / Field Crops Research 101 (2007) 104116

P = 0.05;

**

P = 0.01.

Table 9 Path coefcient analysis showing direct (in bold) and indirect effects of 15 traits over harvest index in Chenopodium Traits Days to Days to Plant owering maturity height (cm) Leaf area (cm2) Branches/ Inorescence Inorescence/ Seed size 1000 seed Dry weight/ Total Leaf Seed Seed Seed plant length (cm) plant (mm) weight (gm) plant (g) chlorophyll carotenoid carotenoid protein Yield (mg/g) (mg/kg) (mg/kg) (%) (t/ha) 0.026 0.056 0.052 0.018 0.065 0.107 0.022 0.021 0.028 0.029 0.005 0.032 0.041 0.028 0.016 0.004 0.006 0.010 0.006 0.009 0.003 0.013 0.003 0.002 0.009 0.007 0.010 0.003 0.002 0.008 0.003 0.019 0.006 0.013 0.009 0.010 0.001 0.049 0.045 0.004 0.007 0.002 0.009 0.001 0.015 0.005 0.009 0.001 0.020 0.006 0.010 0.006 0.033 0.039 0.001 0.015 0.008 0.009 0.004 0.019 0.248 0.346 0.526 0.256 0.423 0.174 0.450 0.055 0.023 0.638 0.191 0.319 0.010 0.276 0.310 0.106 0.030 0.255 0.385 0.069 0.034 0.426 0.095 0.286 0.216 0.722 0.624 0.175 0.055 0.367 0.112 0.346 0.747 0.691 0.575 0.340 0.802 0.049 0.229 0.560 0.966 1.118 0.568 0.254 0.566 0.008 0.022 0.018 0.009 0.027 0.023 0.015 0.010 0.014 0.001 0.015 0.031 0.061 0.004 0.001 0.058 0.102 0.072 0.090 0.093 0.256 0.093 0.219 0.055 0.211 0.065 0.078 0.039 0.309 0.003 0.140 0.028 0.248 0.106 0.246 0.019 0.257 0.056 0.256 0.016 0.004 0.245 0.009 0.004 0.505 Genotypic Correlation 0.070 0.381** 0.410** 0.022 0.367** 0.248 0.237 0.423** 0.426** 0.516** 0.018 0.271* 0.171 0.500** 0.267*

Days to owering 0.169 Days to maturity 0.120 Plant height (cm) 0.083 Leaf area (cm2) 0.057 Branches/plant 0.080 Inorescence length (cm) 0.042 Inorescence/plant 0.056 Seed size (mm) 0.010 1000 seed weight (g) 0.020 Dry weight/plant (g) 0.066 Harvest index 0.025 Total chlorophyll (mg/g) 0.017 Leaf carotenoid (mg/kg) 0.022 Seed carotenoid (mg/kg) 0.040 Seed protein (%) 0.034 *P = 0.05; **P = 0.01.

0.041 0.074 0.073 0.046 0.058 0.092 0.067 0.058 0.035 0.151 0.108 0.083 0.018 0.076 0.215 0.030 0.035 0.128 0.066 0.098 0.030 0.073 0.036 0.059 0.028 0.108 0.091 0.063 0.021 0.016 0.054 0.016 0.013 0.005 0.108 0.014 0.031 0.124 0.086 0.065 0.002 0.053 0.114 0.009 0.018 0.101 0.133 0.050 0.021 0.045 0.031 0.043 0.017 0.058 0.081 0.022 0.010 0.076 0.093 0.041

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path and indirectly and positively inuenced harvest index through most of the traits. 4. Discussion Quinoa is considered a crop adapted to diverse habitats with tremendous potential for diversication of agricultural systems in mountainous regions of the developing world such as the Himalayas and the central mountain region of Africa (Jacobsen, 2001; Jacobsen and Risi, 2001). Quinoa has generated increased interest among farmers, agro-industries and researchers in its native region as well as in North America and Europe. Demand for quinoa has increased considerably during recent years but the supply of quinoa from South American countries is insufcient which necessitates its introduction outside the Andean region. Some new sites have been identied in the American and European Test of quinoa, organized by the FAO. Field trials in Italy and Greece have shown promising results with reported grain yield of 2280 and 3960 kg/ha, respectively (Mujica et al., 2001). Our results show that quinoa could serve as an alternative winter crop for the North Indian Plains and other subtropical regions having similar agro-climatic and edaphic conditions. A thorough assessment of yield potential of all the germplasm lines under study for two consecutive years clearly shows that 41% of the lines were high yielding. This reects greater adaptability of some quinoa lines to North Indian agro climatic conditions, especially of the Chilean and U.S. ones. It was noticed that variability was not related to geographical origin. This is exemplied by the Bolivian lines, two of which (C. quinoa PI 614938 and PI 478408) were the lowest yielding and also recorded low values for most of the traits. However, high seed yield along with above average mean performance for most morphological traits were observed in two other Bolivian lines namely, C. quinoa PI 478414 and C. quinoa Ames 21909. This extreme variation in seed yield witnessed in C. quinoa PI 478408 and C. quinoa PI 478414, which were collected from the same location (La Paz, Bolivia), could be due to genetic factors and points towards wide genetic variation present in the lines having the same origin. Such differences also exist in C. quinoa PI 510536 and C. quinoa PI 510537 whose origin are the same but yield varies by more than six times. One interesting observation was that some lines collected from high elevations (C. quinoa PI 478414, C. quinoa PI 596498, C. quinoa PI 478410 and C. quinoa Ames 21909) gave considerably good seed yield at 120 m altitude of the experimental site. The present results further support earlier reports (Jacobsen, 2003) that quinoa is adapted to areas ranging from sea level to high altitudes. In our study, the US and Chilean lines, and one line each from Argentina (PI 614881) and Bolivia (PI 478414) seem to be adaptable and more suited to subtropical North Indian conditions as they gave consistently high yield, while both lines of C. berlandieri subsp. nuttalliae of Mexican origin were poor in terms of yield. Jacobsen and Stolen (1993) and Jacobsen (2003) reported that Chilean lines have low sensitivity to photoperiod, thus conferring greater stability to them. Our study conrms that

the Chilean lines are more suited for diversication of quinoa in newer areas. These results indicate ample scope for quinoa in furthering agricultural diversication in countries having monsoon climate like India that have markedly cold winters and hot summers. The 29 accessions evaluated had an average pre-owering growth period of about 82 days and took around 48 days for grain maturity. The total growth period in North Indian conditions is less than that reported in South America (110190 days) (Jacobsen and Stolen, 1993) and is similar to that reported in northern Europe (Jacobsen, 1998). The total growth period of quinoa at the experimental site is quite low in comparison to the generally recommended <150 days growing period of quinoa (Jacobsen, 2003). The non-signicant correlation with grain yield and low direct and indirect effects shown by days to owering and days to maturity suggests that these two components are inuencing yield to a lesser extent and are of not much signicance in North Indian conditions. Days to maturity was negatively, though non-signicantly related with total chlorophyll content which was possibly due to gradual rise in temperature and drop in humidity as the season progressed, which leads to degradation of leaf pigments and the enzyme RuBisCo (Garcia del Moral et al., 1995; Fernandez-Figares et al., 2000; Bhargava et al., 2006b). The low yielding accessions generally had below average values for most of the morphological traits and vice versa. This is also corroborated by correlation analysis where all the morphological traits showed positive association with seed yield that was signicant for most of the traits. Thus, different morphological traits seem to positively inuence yield to a great extent. An interesting observation was made with regard to association between days to maturity and seed size, which was negative and highly signicant, thereby indicating a negative impact of maturity period on seed size. Our results are similar to that obtained by Bertero et al. (1999) who reported that temperature and photoperiod after anthesis affects seed diameter to a considerable extent in quinoa. Short photoperiod and cool temperatures after anthesis promote seed diameter, while long photoperiods and high temperature after anthesis negatively affect leaf size (Bertero et al., 1999). Our ndings support their results as owering at the experimental location occurred at the beginning of February, the same time when winter ends. Later on, there is a constant rise in temperature and day length, which progresses till June when the maximum temperatures reach about 45 8C. In our experiment, the control conditions of Bertero et al. (1999) were created naturally as after anthesis the temperature and photoperiod increased which had a negative impact on seed size. Thus, in North India, sowing of quinoa should be done at the onset of winter so that by the time summer approaches, seed formation and development is complete. A strong positive association existed between seed size and 1000 seed weight, which is on expected lines, as a large seed would naturally have more weight than a smaller one. Highly signicant correlations between seed diameter and seed weight have also been determined earlier in quinoa by Ochoa and Peralta (1988), Cayoja (1996) and Rojas et al. (2003). Most of the accessions having above average seed size also had very

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high 1000 seed weight which support the results obtained through correlation. Among other morphological variables, signicant association among branches/plant, inorescence length and inorescence/ plant indicates that plants with good branching habit tend to develop large number of long inorescences. Inorescence length also correlated positively with plant height indicating that lines with greater plant height also developed longer panicles, a fact also reported by Rojas et al. (2003) and Ochoa and Peralta (1988). Biomass accumulation and partitioning to reproductive structures are key determinants of crop yield (Andrade et al., 1999). Reproductive partitioning is the proportion of total biomass allocated to reproductive tissues (Hay, 1995; Sinclair, 1998). The productive capacity of any crop plant depends, not only on its photosynthetic efciency, but also on the effective translocation of assimilates to the seeds, which is measured by the harvest index. This partitioning between vegetative and reproductive parts can be modied by agronomic practices such as sowing density, fertilization, irrigation and choice of sowing date. In the present study, harvest index presented tremendous variability and ranged from 0.26 to 1.43, the highest value being approximately six times the lowest value. However, this range is quite narrow as compared to the report of Rojas et al. (2003) who reported harvest index in quinoa in the range of 0.060.87, a difference of almost 15 times. The harvest index values in the present study are a bit high which might be due to the fact that the plants were cut about 6 cm above the ground and the dry weight excluded the weight of secondary branches and leaves. The inclusion of the above mentioned plant parts in dry weight would in most probability lower the harvest index below 1.00. However, still the harvest index values are pretty high and point towards high efciency of reproductive partitioning in quinoa. Donald (1963) has reported that early sowing, when combined with good conditions and a long growing season leads to severe inter-plant competition and low harvest index. However, our results show no inverse relationship between harvest index and long growing season. Therefore, the variations of sowing date ` and good growing conditions vis-a-vis harvest index in quinoa needs detailed examination. In the present study, days to owering showed positive association with dry weight/plant but none with harvest index. Days to maturity showed strong positive association with dry weight/plant and negative correlation with harvest index, while both dry weight/plant and harvest index were negatively correlated between themselves. Thus, it seems that in the late maturing lines, as season progressed the dry weight increased but there was much less partitioning of the assimilated product to reproductive parts in comparison to the vegetative parts. The present study gave some interesting results with reference to quality traits. A large amount of variation was found with respect to quality traits studied. Earlier, Prakash et al. (1993) have reported signicant differences within quinoa germplasm and suggested their use in breeding of nutritionally superior lines. Carotenoids play an important role in human nutrition as the whole vitamin A comes through diet and many carotenoids have been identied as precursors to

vitamin A while others have been shown to function as antioxidants (de Pee and West, 1996; Rock, 1997; Pavia and Russel, 1999). The leaf carotenoid content was higher than that reported for spinach, amaranth and Chenopodium album (Gupta and Wagle, 1988; Prakash and Pal, 1991; Shukla et al., 2003; Bhargava et al., 2006b, in press). The overall mean for seed carotenoid content was lower in comparison to earlier reports (Koziol, 1992). The total seed protein content corroborates earlier reports by Koziol (1992), Wright et al. (2002) and Repo-Carrasco et al. (2003). The protein content in quinoa is quite high in comparison to commonly used cereals and compares favourably with other underutilized crops like Amaranthus (Bressani et al., 1987; Shukla et al., 2004, 2005), Fagopyrum (Steadman et al., 2001) and even some underutilized legumes like Cassia oribunda (Vadivel and Janardhanan, 2001). The high seed protein in quinoa indicates its potential as a low cost protein source to eliminate protein malnutrition in developing countries where low incomes restricts the use of meat and pulses by the bulk of the population. There is an urgent need for obtaining high quality protein concentrates to solve the problem of chronic malnutrition affecting urban and rural populations in the developing countries. The embryo, after separation from the seed, can be utilized in food for malnutrited children and for pregnant and breast-feeding women. C. quinoa PI 510537 was nutritionally superior, having high protein and carotenoid content coupled with high seed yield. Three germplasm lines of C. quinoa namely CHEN 67/78, CHEN 71/78 and PI 478414 had high carotenoid (both leaf and seed) and protein content but had low seed yield and thus could be used as donor parents for quality improvement. Although top three high yielding accessions had an above average protein, but this relationship does not hold good for other high yielding accessions, which had low protein, therefore, showing insignicant correlation between yield and protein content, which is contrary to the generally accepted view that increase in protein content is made at the expense of yield (Jenner et al., 1991; Pleijel et al., 1999). The non-signicant correlation between seed yield and seed quality traits and low values of direct path are benecial since it would not hinder attempts to breed lines with both greater grain yield and high seed protein and carotenoid. The seed quality traits in quinoa are reported to be negatively inuenced by cold stress and humid autumn weather in high altitude and/or high latitude locations (Jacobsen, 2003). Our results show that at low elevations and low latitude, seed quality traits show no association with duration of growth. This would enable cultivation of quinoa in the tropics and at lower elevations without in any deteriorating the quality of the grain. Protein content is also negatively and indirectly affected by yield through most of the traits, morphological as well as quality. Seed carotenoid seems to be correlated with seed coat colour as six out of the seven dark coloured accessions had high seed carotenoid. Such an association between coat colour and carotenoid content has also been reported in transgenic rice (Datta et al., 2003). Variability plays an important role in any crop-breeding program and determines the limit of selection for yield improvement. The relative amount of genotypic variation is

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best expressed as the genotypic coefcient of variation (GCV), since this variable takes into account the mean value as well as the unit of measurement into consideration. The high GCV for seed yield supports the view that reproductive characters in general are more variable than vegetative ones (Sachs and Coulman, 1983). The heritability values for most of the traits were high suggesting that these traits are under genotypic control. Such high heritability values for various traits have also been reported in vegetable chenopods (C. album) (Bhargava et al., 2003b, 2006b). However, estimation of heritability is of little signicance in coherent selection breeding programs unless accompanied by sufcient genetic gain (Johnson et al., 1955b). Due to large differences in the phenotypic variation between different traits, genetic advance is not directly related to heritability values. In the present study, moderate to high genetic gain values for most of the traits indicate that improvement could be made in the aforesaid characters. The genetic advance for some traits were high because of extreme variation in the material investigated, and smaller values for genetic advance are expected in further selection cycles in a more improved material. Among the various traits, 1000 seed weight, branches/plant, dry weight and total chlorophyll inuenced seed yield to a considerable extent by exhibiting strong positive correlation with seed yield coupled with high positive path and indirect inuence through most of the traits. This indicates scope for improvement in seed yield by proper selection pressure. These traits also showed moderate to high coefcient of variation, heritability and genetic gain values that amply demonstrate the utility of these traits in selection programmes in quinoa. Seed yield and seed protein were the only traits exhibiting high positive direct path and signicant positive association with harvest index, indicating a true relationship among these traits. Leaf carotenoid and dry weight/plant had negative direct effect on harvest index, showed negative signicant correlation and indirectly inuenced harvest index through majority of the traits, indicating actual relationship and suggesting that selection of plants having less dry weight and low leaf carotenoid would likely bring improvement in harvest index. Total chlorophyll had high positive direct effect but exhibited non-signicant negative correlation with harvest index due to presence of strong indirect effect via leaf carotenoid. Thus, the ideotype to increase harvest index in quinoa should have high seed yield and low dry weight/plant. The study of path analysis also indicated that there were no common causal factors that directly inuenced both seed yield and harvest index. Acknowledgements The authors are thankful to the Director N.B.R.I, Lucknow for providing the facilities and constant encouragement to carry out the present investigation. Atul Bhargava acknowledges C.S.I.R., New Delhi for providing nancial assistance. Appendix A. Supplementary data Supplementary data associated with this article can be found, in the online version, at doi:10.1016/j.fcr.2006.10.001.

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