Evaluation of morphological and yield traits in the populations of Vicia spp.

Javadi, Hamideh; Shanjani, Parvin Salehi; Hoseini, Leila Falah; Yeganeh, Masoumeh Ramazani; Javadi, Hamideh; Shanjani, Parvin Salehi; Hoseini, Leila Falah; Yeganeh, Masoumeh Ramazani

doi:10.22319/rmcp.v13i4.6125

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Revista mexicana de ciencias pecuarias

versão On-line ISSN 2448-6698versão impressa ISSN 2007-1124

Rev. mex. de cienc. pecuarias vol.13 no.4 Mérida Out./Dez. 2022 Epub 11-Nov-2022

https://doi.org/10.22319/rmcp.v13i4.6125

Articles

Evaluation of morphological and yield traits in the populations of Vicia spp.

Hamideh Javadi^a^*

Parvin Salehi Shanjani^a

Leila Falah Hoseini^a

Masoumeh Ramazani Yeganeh^a

^{^a} Gene Bank of Research Institute of Forests and Rangelands, Agricultural Research, Education and Extension Organization, Tehran, Iran.

Abstract

The study was focused on estimation of genotypic variation for the morphological and forage yield traits of some vetch genotypes to assess their breeding potential. A small-plot trial was carried out in 2018-2020 at the experimental field of the Research Institute of Forests and Rangelands, Alborz province, Iran. Fifty eight (58) vetch genotypes of Vicia spp. from the natural resources gene bank of Iran, were tested. There was significant (P<0.01) genotypic variation among populations, for all the traits measured. V.monantha (32845) produced high plant and large pods, while V. villosa (322) produced more biomass than other accessions. In the shorter growing seasons, the earliness of V. sativa var.angustifollia (4740,7243), V. sativa var.stenophylla (1862), V. villosa (315, 322) resulted in high seed yield. The principal component analysis showed that the variations observed were mainly caused by traits such as days to flowering and seed ripening and seed traits, that their contribution was important in discriminating the populations. Direct selection can also be made for the populations with high biomass yield based on the recorded performance of these populations during the field experiments. A cluster analysis of the tested vetch populations based on measured traits, at 11.49 genetic distance, created five main groups that showed the similarity of members of each group. Generally, vetch species and their populations had different growth features, phenology, forage and seed productivity. The generated information in this study gives a base for genetics variety of genus Vicia L. and could be useful for including in the future breeding programs.

Key words Biomass yield; Morphological traits; Phenology; Seed yield; Vicia spp

Resumen

El estudio se centró en el cálculo de la variación genotípica para las características morfológicas y de rendimiento de forraje de algunos genotipos de alverja para evaluar su potencial de mejora. En 2018-2020 se llevó a cabo un ensayo en pequeñas parcelas en el campo experimental del Instituto de Investigación de Bosques y Pastizales, provincia de Alborz, Irán. Se analizaron cincuenta y ocho (58) genotipos de alverja (Vicia spp.) del banco de genes de recursos naturales de Irán. Hubo una variación genotípica significativa (P<0.01) entre las poblaciones, para todas las características medidas. V. monantha (32845) produjo una planta alta y de vainas grandes, mientras que V. villosa (322) produjo más biomasa que otras variedades. En las estaciones de crecimiento más cortas, la precocidad de V. sativa var. angustifollia (4740,7243), V. sativa var. stenophylla (1862), V. villosa (315, 322) dio lugar a una elevada producción de semillas. También es posible hacer una selección directa de las poblaciones con un alto rendimiento de biomasa basándose en el rendimiento registrado de estas poblaciones durante los experimentos de campo. Un análisis de conglomerados de las poblaciones de arveja analizadas, basado en las características medidas, con una distancia genética de 11.49, creó cinco grupos principales que mostraron la similitud de los miembros de cada grupo. En general, las especies de arveja y sus poblaciones presentaron características de crecimiento, fenología, productividad de forraje y semillas diferentes. La información generada en este estudio proporciona una base para la variedad genética del género Vicia L. y podría ser útil incluirla en los futuros programas de mejoramiento genético.

Palabras clave Rendimiento de biomasa; Características morfológicas; Fenología; Rendimiento de semillas; Vicia spp

Introduction

Vicia L. is a genus with around 232 species in the world and 45 species in Iran, from the legume family, Fabaceae, as an annual and perennial herb. These species have been known by the common name vetches. The genus is primarily found in the Mediterranean and Irano-Turanian regions, such as in Iran, Anatoly, Caucasus, Iraq, Afghanistan, Central Asia, Talesh, Syria, Armenia, Turkmenistan, Jordan, North Africa, Greece, Pakistan, and Palestine¹. Vetches are short-lived forage plants that are highly resistant to cold and dehydration conditions and can be grown in rainfed and irrigated climates. They fix nitrogen in the soil by fixation in root nodes, and help to soil erosion by planting in sloping areas²^,³. As a legume crop, it provides nitrogen to the soil and reduces the incidence of diseases in succeeding non-leguminous crops. Their widespread adaptation and excellent capacities to produce biomass make them very attractive to farmers⁴. One attraction of vetch is its versatility, which permits diverse utilization as either ruminant feed or green manure. Because of rapid growth in the first year, different species of Vicia spp. can be used to improve overall livestock, feed quality, improve soils, agriculture for fodder, green manure, human nutrition, and the pharmaceutical industry⁵.

Iran is a genetics resources of the genus Vicia and it is widely distributed in different habitats and conditions. Most of the plants in Vicia genus show more variety in morphological traits and sometimes it is difficult to distinguish species of this genus⁶^,⁷.

Genetic variation among Vicia genotypes is imperative for their efficient utilization in plant breeding schemes and effective conservation. Diversity studies available in germplasm, collections have been performed on many plant species for Vicia genus from different regions of the world. In comparison to other annual forage legumes, advances in breeding vetches (Vicia spp.) are rather modest. It has been one of the morphological characteristics of the plant reported in V. sativa⁸^-¹⁴, V. faba¹⁵, V. narbonensis⁸^,¹⁰^,¹¹^,¹⁵, V. ervilia¹⁶, V. villosa¹⁰^,¹¹, V. atropurpurea¹¹, V. dasycarpa⁸, V. hybrid, V. pannonica, V. lutea, V. peregrine, V. lathyroides and V. grandiflora¹¹.

There are 335 accessions of 25 Vicia spp. in natural resources gene bank of Iran, that have been collected from different geographical regions of Iran. In this study, it was aimed to determine some morphological characteristics and forage yields of different vetch genotypes by collecting from natural flora of Iran region. The present study was focused on the estimation of genotypic variation for 12 morphological traits within the V. michauxii, V. michauxii var.stenophylla, V. monantha, V. narbonensis, V. sativa with three varieties: V. sativa var.angustifollia, V. sativa var.cordata, V. sativa var.sativa and V. villosa, to assess their breeding potential and suitability for developing novel common vetch lines with improved agronomic characteristics related to grain production and quality.

Material and methods

Germplasm

A total of 58 germplasm populations were evaluated in this study. This consisted of 1 V. michauxii, 1 V. michauxii var.stenophylla, 1 V. monanta, 1 V. narbonensis, 34 V. sativa, 9 V. sativa var.angustifollia, 1 V. sativa var.cordata, 4 V. sativa var.sativa and 6 V. villosa. The populations were acquired from the Natural Resources Genebank of Iran (Table 1).

Table 1 The list of studied 58 vetch (Vicia spp.) populations

Taxon	Code	Abbre. code	Origin, province	Longitude		Latitude		Altitude (m asl)
*V. michauxii*	2944	Vmi	East Azerbaijan, Kaleybar	47°	02´	38°	51´	1500
V. michauxii var.stenophylla	37129	Vmis	Qom	50°	56´	34°	11´	2482
*V. monantha*	32845	Vmo	Kermanshah	47°	14´	34°	8´	1338
*V. narbonensis*	34878	Vn	Lorestan, Aleshtar	48°	10´	33°	45´	1495
*V. sativa*	5321	Vs	East Azerbaijan	46°	16´	37°	54´	1750
	6646	Vs	Lorestan, Kohdasht	33°	40´	47°	30´	1200
	6654	Vs	Lorestan, Kohdasht	33°	17´	47°	27´	1130
	6681	Vs	Lorestan, Kohdasht	33°	32´	47°	37´	1260
	11760	Vs	Gilan, Rezvanshahr	37°	31´	49°	13´	280
	11761	Vs	Gilan, Rasht	36°	51´	49°	37´	80
	11762	Vs	Gilan, Rezvanshahr	37°	37´	49°	07´	280
	11763	Vs	Gilan, Rasht	37°	59´	49°	33´	100
	11764	Vs	Gilan, Talesh	37°	32´	45°	55´	280
	11771	Vs	Gilan, Talesh	37°	42´	48°	55´	150
	11772	Vs	Gilan, Rezvanshahr	37°	32´	49°	07´	20
	11774	Vs	Gilan, Rasht	37°	11´	49°	39´	120
	24062	Vs	Gilan, Astaneh Ashrafiyyeh	37°	20´	49°	47´	25
	24069	Vs	Gilan, Chabuksar	36°	56´	50°	32´	170
	24074	Vs	Gilan, Astaneh Ashrafiyyeh	37°	19´	50°	07´	16
	24076	Vs	Gilan, Chabuksar	36°	57´	50°	35´	210
	24084	Vs	Gilan, Rahimabad	37°	02´	50°	18´	40
	24097	Vs	Gilan, Rahimabad	37°	01´	50°	17´	45
	32972	Vs	Kermanshah, Hersin	34°	13´	47°	25´	1367
	33456	Vs	Hamadan	47°	57´	34°	24´	1545
	38517	Vs	Gilan, Siyahkal	49°	57´	36°	59´	342
	38523	Vs	Gilan, Talesh	49°	3´	37°	36´	405
	38526	Vs	Gilan	48°	46´	37°	41´	827
	38527	Vs	Gilan, Astra	48°	58´	38°	24´	21
	38528	Vs	Gilan, Rudsar	50°	12´	36°	48´	608
	38531	Vs	Gilan, Rezvan shahr	49°	20´	37°	30´	315
	38532	Vs	Gilan, Talesh	49°	4´	37°	37´	450
	38533	Vs*	Gilan	38°	10´	48°	20´	600
	38536	Vs*	Gilan	36°	54´	49°	26´	577
	40310	Vs	Kermanshah, Salase babajani	34°	49´	46°	05´	1395
	40315	Vs	Kermanshah, Salase babajani	34°	49´	46°	05´	1395
	40326	Vs	Kermanshah, Javanrud	34°	48´	46°	33´	1525
	40334	Vs	Kermanshah, Salase babajani	34°	51´	46°	01´	1395
	43100	Vs	Khozestan, Masjed soliman	31°	56	49°	18´	870
V. sativa var.angustifollia	38524	Vsa	Gilan, Siahkal	50°	14´	36°	53´	670
	38525	Vsa	Gilan, Talesh	48°	51´	37°	41´	281
	38530	Vsa	Gilan, Talesh	48°	52´	37°	41´	215
	38534	Vsa	Gilan, Rasht	49°	35´	37°	0´	137
	38535	Vsa	Gilan, Rodbar	49°	40´	36°	46´	968
	38537	Vsa	Gilan, Gilan	49°	31´	36°	56´	187
	4740	Vsa	Ilam, Ivan	46°	26´	33°	38´	1170
	7243	Vsa	Kohkiloye ve Boyerahmad, Firozabad	52°	57´	28°	86	1900
	38529	Vsa	Gilan, Rezvan shahr	49°	5´	37°	28´	307
V. sativa var.cordata	34295	Vsc	Gilan, Rezvan shahr	49°	4´	37°	36´	310
V. sativa var.sativa	1862	Vss	Kermanshah	47°	06´	34°	31´	1350
	24631	Vss	Kermanshah	47°	06´	34°	31´	1400
	29802	Vss	Kohkiloye ve Boyerahmad	30°	59´	51°	07´	2380
	32900	Vss	Kermanshah	34°	16´	46°	09´	1444
*V. villosa*	315	Vv	Alborz, Karaj	35°	83´	51°	01´	1460
	322	Vv	Karaj	35°	83´	51°	01´	1470
	6268	Vv	Fars, Shiraz, Sepidan, Sheshpir	30°	25´	51°	98´	2350
	14561	Vv	Merkezi, Arak	34°	09´	49°	70´	1730
	28061	Vv	Ardabil	38°	25´	48°	29´	1350
	34212	Vv	Chahar-mahale Bakhtiyari, Borujen	31°	46´	50°	59´	2600

Field trial

Seed of all 58 populations were sown in seedling pots (December 2018). Then planting and maintenance operations were carried out in the field at the research field of Research Institute of Forests and Rangelands, Alborz province, Iran (2018-2020). A week before planting, the soil was prepared as a fine seedbed to enhance good seedling establishment. The field experimental layout was a One-way analysis of variance (ANOVA) designed. The row and plant spacings were 100 and 40 cm, respectively. The trial was managed according to previouse experiences (several hand weeding was practiced, the first hand weeding was made 40 d after crop emergence, and then repeated every forty days until the end of the growing season, to minimize yield reduction due to weed competitions for soil nutrients, water and solar radiation). Irrigation was applied during the trial. The populations were harvested for seed during the period July to November 2020, depending on their maturity.

Morphological traits

Ten plants (normal growth, uniform performance, disease- and insect pest-free) of each 58 Vicia populations were evaluated by 12 different quantitative traits including day to spourat (day to germination), days to first flowering, days to total flowering, days to maturity (days to seed ripening), plant height (at 50 % flowering, cm), internode length (second internode at 50 % flowering, cm), stems number, pod length (cm), pod width (cm), pod index (pod length/width), biomass yield (plant fresh weight) (g), and plant dry weight (g)¹⁷.

Data analysis

Data were subjected to analysis of variance (ANOVA) using the SAS software system¹⁸. Significant differences among the mean values of 12 traits were compared the DMRT Duncan test. Pearson correlation was determined using SPSS v.21. To evaluate the information contained in the collected morphological data, principal component analysis (PCA) was carried out by Minitab software (version 15). PCA was used to identify the most important traits (plant height, internode length, stems number, pod length, pod width, pod index, biomass yield, dry weight, day to sprout, days to first flowering, days to total flowering, days to maturity) in the data set. Mean values populations were used to create a correlation matrix from which the standardized PCA scores were extracted and a Scatter plot on the first two PCA was performed. Cluster analysis was performed using Ward’s methods and Euclidean distance and a dendrogram was calculated.

Results

The results of analysis variance revealed significant (P<0.01) variation for eight morphological and yield traits among taxa and populations of Vicia spp. except for pod width trait among populations (Table 2). Table 3 shows the comparison of mean morphological and yield traits in nine taxa of Vicia spp. The value of plant height, internode length and stems number differ between 24.50-150 cm, 3.29-15 cm and 2.81-9, respectively. The highest value of plant height (150 cm), stems number (9) and internode length (15 cm), were shown in V. monantha (Vmo) and V. michauxii var. stenophylla (Vmis), respectively. The variation of pod length between taxa was significant and it differs from1.06 cm in V. sativa var.cordata (Vsc) to 4 cm in V. monantha (Vmo). There was no significant difference in pod width between taxa and they located as two groups (a and b), so two taxa of V. michauxii (Vmi and Vmis) had the widest pod (1.14 and 1.1 cm). Despite the significant differences in biomass yield and dry weight traits, V. villosa (Vv) showed the most value of these traits (biomass yield=60.12 g and dry weight=15.63 g).

Table 2 Analysis variance of eight morphological traits of 58 vetch (Vicia spp.) populations

Source of Variations	Degrees of freedom (d.f)	Plant height	Internode length	Stem number	Pod length	Pod width	Pod index	Biomass yield	Dry weight
Taxon	8	3770.70**	48.92**	25.93**	5.23**	0.47**	9.19**	3809.63**	223.70**
Population	48	905.59**	12.11**	15.62**	1.89**	0.06 ns	7.76**	1967.42**	202.36**
Error	150	346.70	2.28	2.58	0.39	0.05	1.30	218.67	18.08
CV %		35.88	29.18	32.38	22.77	39.08	22.53	44.56	48.83

*, ** significant at 0.05 and 0.01 levels, respectively; ^ns not significant.

Table 3 Means comparison of 8 traits in different species of Visia spp.

Species	Plant height (cm)	Internode Length (cm)	Stems number	Pop length	Pod width (cm)	Pod index	Biomass Yield (g)	Dry weight (g)
V. michauxii (Vmi)	63.38 ^cd	6.56 ^bc	2.81 ^b	2.54 ^bc	1.14 ^a	4.36 ^b	9.59 ^cd	5.46 ^bc
V. michauxii var. stenophylla (Vmis)	100.0 ^b	15.0 ^a	5.0 ^b	2.5 ^bc	1.1 ^a	2.27 ^c	20.0 ^bd	5.0 ^bc
V. monantha (Vmo)	150.0 ^a	8.0 ^b	9.0 ^a	4.0 ^a	0.6 ^b	6.67 ^a	0.06 ^d	0.01 ^c
V.narbonensis (Vn)	24.50 e	4.75 ^cd	3.0 ^b	3.3 ^ab	0.65 ^b	5.1 ^ab	5.13 ^cd	1.15 ^c
V. sativa (Vs)	48.41 ^de	4.8 ^cd	5.08 ^b	2.88 ^bc	0.56 ^b	5.33 ^ab	35.06 ^b	9.16 ^ab
V. sativa var. angustifollia (Vsa)	45.07 ^de	6.9 ^bc	4.6 ^b	3.27 ^ab	0.60 ^b	5.48 ^ab	19.03 ^bd	5.18 ^bc
V. sativa var. cordata (Vsc)	85.13 ^bc	8.69 ^b	8.75 ^a	1.06 ^d	0.29 ^b	3.75 ^bc	27.92 ^bc	6.92 ^bc
V. sativa var. sativa (Vss)	54.67 ^ce	3.29 ^d	4.58 ^b	2.0 ^cd	0.5 ^b	4.0 ^bc	41.08 ^ab	9.32 ^ab
V. villosa (Vv)	63.56 ^cd	3.36 ^d	4.5 ^b	2.17 ^c	0.51 ^b	4.26 ^b	60.12 ^a	15.63 ^a

Fifty eight (58) populations of Vicia spp. were compared for vegetative and phenology traits (Table 4). There was a wide range of value in plant height from 19 cm in V. sativa var.angustifollia (38534) to 150 cm in V. monantha (32845), also the most value of plant height between populations of species were shown in V. villosa (322) (100.33 cm). V.michuxii var.stenophylla (37129) (100 cm), V. sativa (38527) (90 cm) and V. sativa var.cordata (34295) (85.13 cm). The length of the internode was a very differet from 1.83 cm in V. sativa (24062, 40334, 43100) to 15 cm in V. michauxii var.stenophylla (37129). Also, 9.83, 8.69, and 8.28 cm of internode length were shown in V. sativa (38527), V. sativa var.cordata (34295), and V. sativa var.angustifollia (38525), respectively. The highest and lowest number of stems were 2 and 15, which were shown in two different taxa of V. sativa species (Vsa38530 and Vs11774). This trait in populations of V. villosa was no significant different. Four populations of V. sativa (38527, 33456, 24074 and 32972), V. sativa var.angustifollia(38525) and V. monantha (32845), had the largest pod in terms of length (4-4.53 cm) and populations V. michauxii var.stenophylla (37129), and V. sativa (5321) had the largest pod in terms of width (1.1 and 1.06 cm). In compare of yield traits (biomass yield and dry weight), three populations of V. sativa: Vs11761, Vs24062, Vs40326, and two populations of V.villosa:Vv322, Vv6268, had the most values of these traits. The values of these traits in these populations were Vs11761 (83 and 26 g.), Vs24062 (83 and 26 g), Vs40326 (103.67 and 36.33 g), Vv322 (108.33 and 38.60) and Vv6268 (83.50 and 19.73 g).

Table 4 Means comparison of 12 traits of 58 populations of different species of Vicia spp.

Population	Plant Height (cm)	Internode Length (cm)	Stems Number	Pod Length (cm)	Pod Width (cm)	Pod Index	Biomass Yield (g)	Dry Weight (g)	Day to Sprout	Days to first Flowering	Days to total Flowering	Days to Maturity
Vmi2944	78.56 c-e	7.94 b-e	2.67 g-i	2.87 c-j	0.82 ab	3.72 k-p	10.15 j-n	2.66 i-n	28 a	95 a	115 c	158 c
Vmis37129	100 b	15 a	5 f-i	2.5 f-k	1.1 a	2.27 op	20.00 h-n	5.00 g-n	28 a	95 a	115 c	158 c
Vmo32845	150 a	8 b-e	9 b-d	4 a-c	0.6 bc	6.67 a-h	0.06 n	0.01 n	28 a	95 a	115 c	158 c
Vn34878	24.50 i-k	4.75 f-p	3.00 g-i	3.30 b-g	0.65 bc	5.10 f-n	5.13 l-n	1.16 k-n	28 a	95 a	120 b	162 b
Vs5321	43.88 e-k	4.00 i-p	2.50 hi	2.14 g-l	1.06 a	2.00 p	2.07 mn	0.55 l-n	28 a	95 a	115 c	158 c
Vs6646	44.33 e-k	2.83 n-p	4.67 f-i	2.83 c-j	0.50 bc	5.67 c-k	61.83 b-d	14.61c-f	28 a	95 a	115 c	158 c
Vs6654	28.33 h-k	2.00 p	4.00 f-i	2.00 h-l	0.50 bc	4.00 j-p	42.67 e-i	10.83 d-i	28 a	95 a	115 c	158 c
Vs6681	63.00 c-h	3.33 l-p	5.00 f-i	3.97 a-d	0.50 bc	7.93 ab	74.83 bc	18.43 cd	28 a	95 a	115 c	158 c
Vs11760	56.67 c-k	4.50 g-p	4.67 f-i	2.00 h-l	0.50 bc	4.00 j-p	53.00 c-g	15.27 c-f	28 a	95 a	115 c	158 c
Vs11761	66.67 b-g	6.17 c-l	5.33 f-h	2.50 f-k	0.50 bc	5.00 f-n	83.00 ab	26.00 b	28 a	95 a	115 c	162 b
Vs11762	37.67 f-k	3.67 j-p	6.00 e-g	2.50 f-k	0.50 bc	5.00 f-n	19.00 h-n	5.06 g-n	28 a	95 a	115 c	158 c
Vs11763	80.00 b-e	6.33 c-i	11 bc	3.33 b-g	0.50 bc	6.67 a-h	45.00 e-h	9.33 e-k	28 a	95 a	120 b	162 b
Vs11764	61.67 c-i	5.17 e-o	11.67 b	2.67 e-k	0.50 bc	5.33 e-m	51.17 c-g	9.03 e-l	28 a	95 a	115 c	158 c
Vs11771	52.33 d-k	4.83 f-p	4.33 f-i	2.00 f-k	0.50 bc	4.00 j-p	56.67 b-f	11.43 d-h	28 a	95 a	115 c	158 c
Vs11772	65.00 b-h	3.17 m-p	4.67 f-i	2.00 f-k	0.50 bc	4.00 j-p	41.50 e-i	8.93 e-l	28 a	95 a	120 b	162 b
Vs11774	68.33 b-f	4.33 h-p	15.00 a	2.67 e-k	0.50 bc	5.33 e-m	53.00 c-g	15.27 c-f	28 a	95 a	115 c	158 c
Vs24062	40.00 f-k	1.83 p	4.67 f-i	2.00 f-k	0.50 bc	4.00 j-p	83.00 ab	26.00 b	28 a	95 a	120 b	162 b
Vs24069	41.00 f-k	2.50 n-p	5.33 f-h	3.83 a-e	0.50 bc	7.67 a-d	37.17 e-k	10.43 d-j	28 a	95 a	115 c	158 c
Vs24074	49.67 d-k	4.17 h-p	6.00 e-g	4.00 a-c	0.47 bc	8.67 a	44.00 d-h	10.43 d-j	28 a	95 a	115 c	158 c
Vs24076	41.33 f-k	3.50 k-p	4.33 f-i	2.50 f-k	0.47 bc	5.50 d-l	41.50 e-i	8.93 e-l	28 a	95 a	120 b	162 b
Vs24084	55.00 c-k	5.33 d-o	4.67 f-i	2.67 e-k	0.50 bc	5.33 e-m	44.00 d-h	10.43 d-j	28 a	95 a	115 c	158 c
Vs24097	51.33 d-k	3.00 n-p	6.00 e-g	2.00 f-k	0.50 bc	4.00 j-p	45.00 e-h	8.77 e-m	28 a	95 a	115 c	158 c
Vs32972	44.67 e-k	2.83 n-p	3.00 g-i	4.00 a-c	0.57 bc	7.11 a-f	48.00 c-h	15.10 c-f	28 a	95 a	120 b	162 b
Vs33456	43.43 e-k	6.07 c-m	4.86 f-i	4.04 a-c	0.64 bc	6.39 b-i	21.29 h-n	5.30 g-n	28 a	95 a	115 c	158 c
Vs38517	34.67 f-k	6.17 c-l	3.33 f-i	2.50 f-k	0.53 bc	4.72 g-n	11.27 j-n	3.05 h-n	28 a	95 a	115 c	158 c
Vs38523	43.33 e-k	7.33 c-g	3.67 f-i	3.17 b-h	0.60 bc	5.28 f-m	25.87 g-n	7.17 f-n	28 a	95 a	115 c	158 c
Vs38526	86.17 b-d	6.50 c-j	6.33 f-i	2.73 e-k	0.40 bc	6.18 b-j	18.92 h-n	5.50 g-n	28 a	95 a	115 c	158 c
Vs38527	90.00 bc	9.83 b	3.00 g-i	4.53 a	0.60 bc	7.56 a-e	20.41 h-n	3.91 h-n	28 a	95 a	115 c	162 b
Vs38528	53.40 c-k	8.00 b-e	6.40 d-f	3.63 a-f	0.63 bc	5.75 b-k	34.49 e-l	8.91 e-l	28 a	95 a	115 c	158 c
Vs38531	46.00 e-k	6.40 c-k	5.20 f-i	2.84 c-j	0.60 bc	4.68 h-n	13.00 i-n	3.95 h-n	28 a	95 a	115 c	158 c
Vs38532	29.80 g-k	7.60 b-f	4.80 f-i	2.00 f-k	0.64 bc	3.27 l-p	8.35 k-n	2.00 i-n	28 a	95 a	115 c	162 b
Vs38533	33.30 f-k	4.74 f-p	3.40 f-i	3.66 a-f	0.62 bc	5.93 b-k	10.54 j-n	2.29 i-n	28 a	95 a	115 c	158 c
Vs38536	36.33 f-k	7.00 c-h	3.67 f-i	2.23 g-k	0.50 bc	4.47 h-o	3.04 mn	0.83 k-n	28 a	95 a	115 c	158 c
Vs40310	33.33 f-k	3.17 m-p	3.67 f-i	3.33 b-g	0.50 bc	6.67 a-h	55.00 c-g	15.77 c-e	28 a	95 a	115 c	158 c
Vs40315	38.33 f-k	2.50 n-p	3.67 f-i	3.17 b-h	0.50 bc	6.33 b-i	51.33 c-g	14.63 c-f	28 a	95 a	115 c	158 c
Vs40326	44.67 e-k	2.33 op	4.33 f-i	3.50 a-f	0.50 bc	7.00 a-f	103.67 a	36.33 a	28 a	95 a	120 b	162 b
Vs40334	31.33 f-k	1.83 p	4.67 f-i	3.33 b-g	0.43 bc	7.83 a-c	41.50 e-i	8.93 e-l	28 a	95 a	115 c	158 c
Vs43100	22.00 j-k	1.83 p	4.33 f-i	1.50 kl	0.50 bc	3.00 n-p	9.57 j-n	0.33 mn	28 a	95 a	120 b	162 b
Vsa38524	29.60 g-k	7.70 b-f	4.40 f-i	3.16 b-h	0.78 ab	4.07 j-p	7.18 l-n	1.95 j-n	28 a	95 a	115 c	158 c
Vsa38525	50.44 d-k	8.28 b-d	5.56 f-h	4.22 ab	0.61 bc	6.96 a-g	19.92 h-n	5.62 g-n	28 a	95 a	115 c	162 b
Vsa38530	65.33 b-h	6.67 c-i	2.00 i	3.17 b-h	0.53 bc	5.69 c-k	29.33 e-n	10.98 d-h	28 a	95 a	115 c	162 b
Vsa38534	19.00 k	5.50 d-n	5.00 f-i	3.25 b-h	0.50 bc	6.50 b-i	26.75 f-n	5.80 g-n	28 a	95 a	115 c	162 b
Vsa38535	36.50 f-k	6.50 c-j	2.50 hi	2.75 d-k	0.65 bc	4.29 i-o	4.55 l-n	1.25 k-n	28 a	95 a	115 c	158 c
Vsa38537	43.67 e-k	8.00 b-e	5.67 f-h	2.93 c-i	0.60 bc	4.89 f-n	4.70 l-n	1.05 k-n	28 a	95 a	115 c	162 b
Vsa4740	49.67 d-k	3.50 k-p	5.00 f-i	2.00 h-l	0.50 bc	4.00 j-p	45.00 f-h	9.33 e-k	21 b	90 b	107 d	150 d
Vsa7243	54.33 c-k	5.17 e-o	4.33 f-i	2.67 e-k	0.50 bc	5.33 e-m	18.67 h-n	5.67 g-n	21 b	90 b	107 d	150 d
Vsa38529	30 f-k	4 i-p	3 g-i	0.6 m	0.3 c	2.00 p	3.10 mn	0.60 l-n	28 a	95 a	115 c	158 c
Vsc34295	85.13 b-d	8.69 bc	8.75 c-e	1.06 l	0.29 c	3.75 k-p	27.92 e-n	6.92 f-n	28 a	95 a	115 c	158 c
Vss1862	44.67 e-k	4.33 h-p	4.33 f-i	2.00	0.50 bc	4.00 j-p	31.67 e-m	9.00 e-l	21 b	90 b	107 d	150 d
Vss24631	51.00 d-k	2.33 op	5.00 f-i	1.83 i-l	0.50 bc	3.67 k-p	56.67 b-f	11.43 d-h	21 b	90 b	125 a	167 a
Vss29802	67.33 b-g	3.50 k-p	4.33 f-i	2.50 h-l	0.50 bc	5.00 f-n	41.50 e-i	8.93 e-l	28 a	95 a	107 d	150 d
Vss32900	55.67 c-k	3.00 n-p	4.67 f-i	1.67 j-l	0.50 bc	3.33 l-p	34.50 e-l	7.93 e-n	28 a	95 a	107 d	150 d
Vv315	55.00 c-k	2.83 n-p	4.33 f-i	2.00 h-l	0.50 bc	4.00 j-p	53.83 c-g	13.08 c-g	21 b	90 b	107 d	150 d
Vv322	100.33 b	4.00 i-p	4.33 f-i	2.50 f-k	0.50 bc	5.00 f-n	108.33 a	38.60 a	28 a	95 a	115 c	158 c
Vv6268	47.67 e-k	3.17 m-p	4.67 f-i	1.67 j-l	0.53 bc	3.17 m-p	83.50 ab	19.73 bc	21 b	90 b	107 d	150 d
Vv14561	58.33 c-j	4.50 g-p	4.67 f-i	2.67 e-k	0.50 bc	5.33 f-n	57.71 b-e	10.83 d-i	28 a	95 a	115 c	158 c
Vv28061	51.67 d-k	2.00 p	4.33 f-i	2.00 h-l	0.50 bc	4.00 j-p	18.00 h-n	3.73 h-n	28 a	95 a	115 c	158 c
Vv34212	68.33 b-f	3.67 j-p	4.67 f-i	2.17 g-l	0.53 bc	4.06 j-p	39.33 e-i	7.80 e-n	28 a	95 a	115 c	158 c

Different letters indicate signiﬁcant differences among different populations for the same species. P <0.05.

V. michauxii (Vmi), V. michauxii var. stenophylla (Vmis), V. monantha (Vmo), V.narbonensis (Vn), V. sativa (Vs), V. sativa var. angustifollia (Vsa), V. sativa var. cordata (Vsc), V. sativa var. sativa (Vss), V. villosa (Vv).

The results of phenology traits showed that all of the populations based on day to sprout and days to first flowering traits were divided into two groups (a and b). V. narbonesis (34878), V. monantha (32845) and two taxa of V. michauxii (Vmi2944 and Vmis37129) had the same value in day to sprout and days to first flowering traits, but populations V. sativa var.angustifollia (Vsa4740, Vsa7243), V. sativa var.stenophylla (Vss1862, Vss24631) and two populations of V. villosa (Vv315, Vv6268) with 21 and 90 d for day to sprout and first flowering were separated from the rest of populations by earlier germination and flowering. In days to total flowering and maturity traits, populations were divided as four groups (a, b, c and d). Days to total flowering as four groups:125a,120b,115c,107d and seed maturity:167a,162b,158c,150d. Populations in group d (107 and 150 d of flowering and seed maturation) had the shortest time required for full flowering and seed maturation. That is, they reached full flowering and seed maturity earlier than other populations. Populations of V. sativa var.angustifollia (Vsa4740, Vsa7243), V. sativa var.stenophylla (Vss1862, Vss29802, Vss32900) and V. villosa (Vv315, Vv6268), having the shortest day for full flowering and seed maturation (Table 4).

Analysis of the genetic correlations among the mentioned traits in the tested vetch populations revealed the existence of several significant positive coefficients (Table 5), namely between plant height with internode length (r_gxy=0.43; P<0.01), stems number (r_gxy=0.38; P<0.01) and pod length with internode length (r_gxy=0.24; P<0.05), pod width (r_gxy =0.23; P<0.05), day to sprout (r_gxy=0.28; P<0.05), days to first flowering (r_gxy=0.28; P<0.05) and days to maturity (r_gxy=0.26; P<0.05), pod index with day to sprout (r_gxy =0.23; P<0.05) and days to first flowering (r_gxy=0.23; P<0.05). On the other hand, the relationship between pod width with pod index (r_gxy =−0.26; P<0.05), biomass yield (r_gxy =−0.35; P<0.01), and dry weigth (r_gxy =−0.28; P<0.05), internode length with dry weigth (r_gxy=−0.38; P<0.01) were negatively and significant.

Table 5 Simple correlation matrix for the 12 traits of Vicia spp. populations

Traits	Plant height	internode length	Stems number	Pod length	Pod width	Pod index	Biomass yield	Dry weight	Day to sprout	Days to first flowering	Days to total
Internode length	0.43**
Stems number	0.38**	0.11^ns
Pod length	0.13 ^ns	0.24*	0.03 ^ns
Pod width	0.10 ^ns	0.51 ^ns	-0.22 ^ns	0.23*
Pod index	0.09 ^ns	-0.06 ^ns	0.16 ^ns	0.86 ^ns	-0.26*
Biomass yield	0.11 ^ns	-0.46 ^ns	0.14 ^ns	-0.03 ^ns	-0.35**	0.20 ^ns
Dry weight	0.13 ^ns	-0.38**	0.07 ^ns	0.03 ^ns	-0.28*	0.22 ^ns	0.95 ^ns
Day to sprout	0.04 ^ns	0.19 ^ns	0.06 ^ns	0.28*	0.11 ^ns	0.23*	-0.16 ^ns	-0.09 ^ns
Days to first flowering	0.04 ^ns	0.19 ^ns	0.06 ^ns	0.28*	0.11 ^ns	0.23*	-0.16 ^ns	-0.09 ^ns	1 ^ns
Days to total flowering	-0.08 ^ns	-0.03 ^ns	0.05 ^ns	0.17 ^ns	0.03 ^ns	0.15 ^ns	0.04 ^ns	0.07 ^ns	0.46 ^ns	0.46 ^ns
Days to maturity	-0.07 ^ns	0.13 ^ns	0.02 ^ns	0.26*	0.06 ^ns	0.21 ^ns	-0.03 ^ns	0.02 ^ns	0.50 ^ns	0.50 ^ns	0.92 ^ns

*, ** significant at 0.05 and 0.01 levels, respectively; ^ns not significant.

Two-dimensional principal component analysis showing the relationship among quantitative traits of studied populations is presented in Figure 1. Populations V. sativa var.angustifollia (4770, 7243), V. sativa var.sativa (1862), V. villosa (315, 6268) were separated partially by PC1; traits related to this separation are mainly phenology traits (day to spourat, days to first flowering, days to total flowering, days to maturity).

Figure 1 Two principal components showing the relationship among 12 traits of 58 populations of Vicia spp.

A cluster analysis of the tested Vicia spp. populations showed five main groups (Table 6 and Figure 2). Cluster G₁ contained five populations, belonging to V.sativa var.angustifollia with two populations (7243, 4740), V. sativa var.sativa one population (1862) and V. villosa with two populations (315, 6268).They are characterized by the lowest values of phenology traits (day to spourat, days to first flowering, total flowering, and seed maturity). Cluster G₂ contained 13 populations: 11 populations belonging to V. sativa (6646, 6681, 11761, 24062, 24069, 24074, 32972, 40310, 40315, 40326, 40334), population 38530 of V. sativa var.angustifollia and 322 of V. villosa. They are also characterized by the highest amount of vegetative, seed and yield traits compared to other populations. Cluster G₃ included 16 popullations belonging to V. sativa (6654, 11760, 11762, 11771, 11772, 24076, 24084, 24097, 43100), population 38529 of V. sativa var.angustifollia, V. sativa var.sativa (24631, 29802, 32900), 28061, 34212 and 14561 of V. villosa, with high amount of vegetative traits were collected in one group. Cluster G₄ contained seven populations: five belong to species V. sativa (11763, 11764, 11774, 38526, 38527), population 34295 of V. sativa var.cordata and V. monantha (32845). These were classified with the highest plant height, stems number and vegetative traits compared with other clusters. Cluster G₅ was the largest one with 17 populations, nine from V. sativa (5321, 33456, 38517, 38523, 38528, 38531, 38532, 38533, 38536), five from V. sativa var.angustifollia (38524, 38525, 38534, 38535, 38537), V. michauxii (2944), V. michauxii var.stenophylla (37129) and V. narbonensis (34878).These were classified as highest vegetative and pod traits populations.

Table 6 Means comparison of 12 traits of five vetch groups produced in Figure 2

Groups	Plant height (cm)	internode length (cm)	Stems number	Pod length (cm)	Pod width (cm)	Pod index	Biomass yield (g)	Dry weight (g)	Day to sprout	Days to first flowering	Days to total flowering	Days to maturity
G₁	50.27 ^b	3.80 ^c	4.53 ^c	2.07 ^c	0.51 ^b	4.10 d	46.53 ^b	11.36 ^b	21.00 ^c	90.00 ^c	107.00 ^c	150.00 ^c
G₂	50.97 ^b	3.40 ^c	4.36 ^c	3.24 ^a	0.50 ^b	6.51 ^a	63.15 ^a	18.94 ^a	28.00 ^a	95.00 ^a	116.15 ^a	159.54 ^a
G₃	49.50 ^b	3.43 ^c	4.60 ^b	2.04 ^c	0.49 ^c	4.14 d	37.73 ^c	8.20 ^c	27.56 ^b	94.69 ^b	115.56 ^b	158.31 ^b
G₄	88.76 ^a	6.98 ^b	9.25 ^a	3.00 ^b	0.48 ^c	5.93 ^b	30.92 ^c	7.14 ^c	28.00 ^a	95.00 ^a	115.71 ^b	159.14 ^a
G₅	43.91 ^c	7.12 ^a	4.21 ^c	3.01 ^b	0.68 ^a	4.72 ^c	13.43 d	3.44 d	28.00 ^a	95.00 ^a	115.29 ^b	159.18 ^a

^abc Different letters indicate signiﬁcant differences among different populations for the same species. P<0.05.

Figure 2 Dendrogram of 58 populations of Vicia spp. explained by complete linkage clustering of 12 traits

The principal component analysis (PCA) of the 12 quantitative traits is summarized in Table 7. The first five PCs had eigenvalues >1 and they explained more than 80 % of the total variation for the vegetative and phenology traits. Day to sprout, days to first flowering, days to total flowering and days to maturity were loaded highly in PC1 and they accounted for 25.7 % of the total variation. In PC2, Biomass yield and dry weight accounted for 21 % of the total variation. In PC3, plant height and internode length accounted for 14.3 % of the total variation. PC4 contributed 11.2 % of the total traits variation in these populations with plant length and stems number loading highly. PC5 accounted for 9.8 % of total variation with length, width and pod index. Generally, for the 12 vegetative and phenology traits studied, PC1 and PC2 constituted more than 46 % of the total traits variation with most phenology traits and yield-related traits. This indicated that these traits can be used to classify the populations under study.

Table 7 Eigenvalues, the proportion of variance, and morphological traits that contributed to the first five principal components (PC)

Variable	PC1	PC2	PC3	PC4	PC5
Plant height	0.058	-0.029	0.466	0.441	-0.267
Internode length	0.208	0.302	0.428	0.055	-0.242
Stems number	0.050	-0.107	0.176	0.668	0.075
Pod length	0.310	-0.154	0.389	-0.244	0.378
Pod width	0.141	0.303	0.243	-0.301	-0.357
Pod index	0.230	-0.332	0.265	-0.084	0.533
Biomass yield	-0.144	-0.53	-0.004	0.062	-0.236
Dry weight	-0.101	-0.541	0.043	-0.042	-0.321
Day to sprout	0.464	-0.017	-0.102	0.045	-0.075
Days to first flowering	0.464	-0.017	-0.102	0.045	-0.075
Days to total flowering	0.380	-0.129	-0.338	0.057	-0.164
Days to maturity	0.419	-0.100	-0.267	0.014	-0.151
Eigenvalue	3.340	2.723	1.856	1.452	1.279
Proportion	0.257	0.210	0.143	0.112	0.098
Cumulative	0.257	0.467	0.610	0.721	0.820

Discussion

In these study, 58 populations of Vicia spp. were investigated for genetic diversity based on morphological and phenology traits. Due to, genetic diversity analysis of germplasms using morphological traits is an initial step for crop improvement¹⁹^-²². There was significant (P<0.01) genotypic variation among 58 germplasm accessions of Vicia spp. for all the measured vegetative and yield traits: plant length, internode length, stems number, pod length, pod width, pod index, biomass yield and dry weight.The estimates of genotypic variation and repeatability for these traits indicated the potential genetic variation available among the germplasm accessions within Vicia spp. investigated. Similar results were obtained by the Ebrahimi et al²³ on plant and seed morphology traits of white Bean genotypes, Mikic et al¹² on forage and seed yields of three lines of common vetch and Berhanu and Abera²⁴ on forage yield of vetch species investigation.

A comparison between taxon (V. sativa: Vs, Vsa, Vsc and Vss, V. mchauxii: Vmi and Vmis, V. monantha: Vmo, V. narbonensis: Vn and V. villosa: Vv) showed V. monantha (Vmo) with high values of plant height, stems number, pod length and V. villosa with high values of biomass yield and dry weight. Berhanu and Abera²⁴ showed that among the vetch species (V. sativa, V. villosa, V. dasycarpa, and V. bengalensis), V. dasycarpa and V. villosa were the best performing species for forage. Then the vetch species tested in the current study could be used for pasture expansion and forage production, in livestock exclusion areas, in forage strips, as an under-sowing with food crops, or as a backyard forage crop in the pasture of the country.

The populations demonstrated high variation in plant height, internode length, stem number, pod length, biomass yield and dry weight. Populations: Vmo32845, Vv322, Vmis37129 (for plant height), populations: Vmi37129, Vs38527, Vsc34295 (for internode length), populations: Vs11774,Vs11764,Vs11763 (for stems number), populations: Vs38527, Vmo32845, Vsa38525 (for pod length) and populations:Vs11761, Vs24062, Vs40326, Vv322, Vv6268 (for biomass yield and dry weight) showed the highest values of the mentioned traits. However, broadening the genetic base from diverse sources is recommended to include most of the genetic determinants of these traits²⁵. This variability can be exploited in fodder breeding programs to select an adapted plant material for the arid and semi-arid areas²⁶.

Phenology (earliness and lateness) of vetch species has a great effect on seed yield productivity. Late maturity for forage and seed was recorded at 125 and 167 d, respectively. This could be due to high and extended rainfall at the region of populations that encouraged vegetative growth and delayed forage and seed harvesting stages. The results indicated that for vetch populations tested, 107 to 125 and 150 to 167 d were required after the emergence of the seedlings for total flowering and seed maturity, respectively. On average, the difference in harvest forage and seed yield between populations are about 18 and 17 d. This indicates different responses of the tested populations for these important agronomic traits.

According to Getnet et al²⁷, Vicia narbonensis and Vicia sativa are early maturing and Vicia villosa is late maturing species. But in this study two populations of V. villosa (315 and 6268) and four populations of V.sativa var.angustifollia (4740, 7243), V. sativa var.sativa (1862, 29802, 32900) with 107 and 150 d for flowering and seed maturity is recommended for seed production due to earliness, whereas late-maturing species like V. sativa var.sativa (24631) should not be advisable to grow for seed purpose.

There is a direct relationship between plant length with internode length and the number of stems, this indicates that tall plants produce long internodes and more number stems. Also, the length and width of the pod have a direct relationship with the number of days of sprout, flowering and seed maturation, which means long and wide pods are produced by late-flowering and seed maturation time. Since, in cereals, the correlation between grain yield and plant height is often negative, but in legumes, this correlation is often positive, because legumes have unlimited growth, therefore, with increasing height, more pods are produced, which has a positive effect on performance, so similar results were obtained in the traits of white Bean genotypes where high grain yields were strongly correlated to days to flowering and plant height²³ and Lens spp.²⁸.

In PCA, since the first component includes changes that are not explained by the second component and the two components are independent of each other, so the two components were intersected vertically and in the form of a biplot diagram to determine the diversity between different genotypes and determine the far and near genotypes to be used. Phenology traits (day to sprout, days to first flowering, days to total flowering, days to maturity) accounted for the variations recorded in the populations in PC1. On the other hand, yield traits (biomass yield and dry weight) accounted for the variation observed in the populations in PC2. The total cumulative variance in the first two PC was more than 46 %, indicating the high degree of diversity among the traits under study. Furthermore, the traits can be used as phenotypic traits in differentiating the populations. In plot PCA (Figure 1), populations, V. sativa var.angustifollia (Vsa7243, Vsa4740), V. sativa var.sativa (1862) and V. villosa (Vv315, Vv6268), separated from other populations and located on the left of X-axis by containing less of phenology traits (important in the first component). So, these populations recommend for areas with short growth periods. Populations V. sativa (40326) and V. villosa (322), for containing high value of biomass and dry weight, located on the bottom of Y-axis (negative effect of biomass and dry weight on the second component). As a result, two populations, V. sativa (40326) and V. villosa (322), produce forage yield more than other populations.

In the present study, the 58 populations of Vicia spp. were grouped into five clusters using 12 traits.The populations of cluster G₁ are characterized by the lowest values of days to sprout, flowering, and seed maturity which are the candidate of further evaluations. Also, these populations had a shoter time for these traits. Members of G₁ are similar to the dispersion of these populations in the PCA plot (Figure 1). It is interesting that the population from different climates like Shiraz clustered with populations from Karaj. This pattern of clustering indicates, the diversity of populations within these geographical areas and, the similarity of populations from different geographical areas.

These results agree with the report of Alemayehu and Becker²⁹ in Brassica carinata. Cluster G₂ contained 13 populations belonging to V. sativa and V. villosa species. These populations had a high value of seed, yield and phenology traits. Member of G₂ due to having a long time for flowering and seed maturity, produce more seed and forage yield. This is the best factor, that can be used for livestock feeding. Cluster G₃ contained mixed 16 populations of V. sativa and V. villosa. with lowest values of seed and forage yield gather together in a group, that they are not important inbreeding. Cluster G₄ contained seven populations of V. sativa and V. monantha with high vegetative traits, that recommend livestock feeding and control of erosion. G₅ group with 17 populations of V. sativa, V. michauxii and V. narbonensis were classified as later flowering and seed ripening and containing less amount of yield forage. These populations can be used for areas with a long growth time.

Finally in this study populations were located as five groups based on morphology and phenology traits. Members of each group are similar for mentioned traits and can be recommended for breeding programs. Also, the results indicated no relationship between studied traits and the origin of populations.

Conclusions and implications

The findings showed the high variation of morphology and yield traits in different species and populations of vetch. These differences are very important to select the type of companion crops and methods of integration to improve yields of both crops (food and forage) without significant effect of one on the other. Vicia sativa (Kermanshah, Javanrod) and V. villosa (Karaj) were superior in terms of fresh and dry forage yields. V.michauxii var.stenophylla (Qom), V. monantha (Kermanshah), V. sativa (Gilan, Astara), and V. villosa (Karaj), are recommended by having tall plant and big pods. However, more comprehensive studies and additional experiments are required to complete information for breeding programs.

Acknowledgments

The authors thank Dr. Jalilian for identification of plants and the director of Gene Bank for providing seeds and making the lab facilities available for our study and RIFR in Iran for financial support.

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Received: December 22, 2021; Accepted: April 21, 2022

^*Corresponding author: Hjavadim@yahoo.com; Javadi@rifr-ac.ir

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