Indian Journal of Agricultural Research

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Modification in Physiological Performances of French Bean (Phaseolus vulgaris L.) Seed Through Seed Priming

Bingiala Laloo2, Prabir Chakraborti1
1Department of Seed Science and Technology, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia-741 252, West Bengal, India.
2School of Crop Improvement, College of PG Studies, Central Agricultural University (Imphal), Umiam-793 103, Meghalaya, India.
Background: The three priming modes using various organic and inorganic compounds for hydro-priming, osmo-priming (PEG 6000, CaCl2, KNO3) and halo-priming (thiourea, GA3) with control were considered, to study the improvement in physiological performances of French bean seed. 

Methods: The seeds were primed for 6 hours duration and observations associated to seven parameters considering seedling and biochemical activities were considered for getting the real exposure in physiological performances of seed for seedling establishment.

Result: The treatment, T2 (thiourea, 750 ppm) was best performing with significant variation among diverse treatments except in peroxidase activity. In genotypes, V3 (RCM-FB-62) showed high significant demarcation for maximum parameters though, V2 (phyrngop) and V1 (RCM-FB-18) were highest for peroxidase activity and speed of germination. The treatment-genotype interactions showed significant demarcation with exemption in speed of germination and vigour index, however, T2V3 interaction was highest considering all parameters. Considerable parameters were pronounced contributors for cumulative upgradation in physiological performances of seed with exceptions in percent of germination and peroxidase activity. Diverse treatments specified the extent of effects on seedling or enzyme activity to upgrade the quality. Consequently, the treatment T2 resulted in improved physiological performance in French bean seed production.
French bean is an important cool season (15-20°C) annual legume crop which is popular for human consumption for its utilization as both eatable dry seed and unripe fruit (green beans). In India, French bean production is emerging due to its adequacy in communities, simple cultivation methodology and distinct dietary status. The high nourishing value of French bean comprises 17.5-28.7 per cent protein in the dry seeds and about 1.0-2.5 per cent protein in the green pods, about 61.4 per cent carbohydrates, 3.2-5.0 per cent mineral matter, 340-450 kcal energy and about 11 per cent water (Fabbri and Crosby, 2016) although, it may vary from region to region due to developmental variation in plant or seed. The wild forms of small black seeds are found in Tropical America from where it was disseminated to West Africa and later to India. The various genotypes have been categorized into ‘pole beans’ and ‘bush beans’ according to its differential climbing habit. The production and dissemination of quality seed is one of the indispensable factors to achieve anticipated yields in addition to optimum quality of the cultivar. The quality of seed exclusively, is responsible for at least 10-15% rise in productivity (Monalisa et al., 2017). The utilization of quality seeds is one of the prime factors that can minimize the vast yield gap between research and farmers’ plot.
Seed priming is one of the invigoration techniques where germination procedure is commenced but appearance of radicle is arrested. Presently, this technology is used for the different crops predominantly in horticultural crops, but crop specific modes of priming need to be developed to further enhance seed production. Seed priming gives rise to greater percentage of final germination, where the seedling parameters, speed of germination are extensively improved. Moreover, a promising reflection specified the decline of dormancy with upgradation of seedling quality (Anese, 2011). The earliness and uniformity in seed germination is habitually attained in application of priming which is vital for establishing a good crop stand (Khan et al., 2008). The biochemical activities required to initiate the germination process were also enhanced as a result of priming (Afzal et al., 2011). The procedure of priming can be beneficial to farmers because it is used as safeguards escaping additional cultivation time, fertilizer applications, re-seeding, weak plants in addition to money. On-farm seed priming can be carried out amongst cooperating farmers because it is simple, cheap and additionally effective for qualitative progression of seedling. Lentil seeds primed with hydro or osmo-priming (KNO3 or PEG-6000) showed hopeful outcomes in seedling performances (Ghassemi-Golezani et al., 2008 and Saglam et al., 2010).
The experiment was conducted at 2016 and 2017 in RKVY Laboratory, Deptt. of Seed Science and Technology, Bidhan Chandra Krishi Viswavidyalaya, West Bengal using six months old stored seed under ambient condition considering three (3) French bean genotypes namely RCM-FB-18 (V1), Phyrngop (V2), RCM-FB-62 (V3). The twelve (12) varied priming treatments comprised of various organic and inorganic chemicals viz. T1 (500 ppm), T2 (750 ppm Thiourea), T3 (50 ppm Gibberellic Acid), T4 (100 ppm Gibberellic acid), T5 (0.1 Mpa KNO3), T6 (0.2 Mpa KNO3), T7 (0.1 Mpa CaCl2), T8 (0.2 Mpa CaCl2), T9 (0.1 Mpa PEG 6000), T10 (0.2 Mpa PEG 6000), T11 (Distilled Water), T12 (Control, without treatment). The dissimilar varietal seeds were soaked separately for 6 hours in aqueous solutions of these treatments followed by drying back to the original seed moisture content, except control. The five seedling parameters (germination percentage, speed of germination, seedling length, seedling dry weight and vigour index) were projected through Glass-Plate method (Chakraborti, 2010). The effect of priming on two important enzymes (alpha amylase and peroxidase) after 24 hours of imbibition was also studied. The assessment was done using OPSTAT software following CRD of two factors with three replications for each character. The outcome was achieved at one percent level of significance.
Considering the diverse seed priming with 12 treatments (T1-T12) including control (without treatment), the observations were related to physiological performances of seed on three genotypes (V1, V2 and V3) of French bean. The explanations were regulated to facilitate the seed quality associated to seedling parameters and biochemical activity in two successive years and pooled value. The various treatments showed significant variances for all considerable parameters which were clustered into two different kinds like seedling parameters and biochemical parameters. In application of diverse priming mode, the variable significant distinction showed advancement, similar to observation of other researchers (Ghobadi et al., 2012 and Abnavi et al., 2012) predominantly the pooled values. The superior treatment values over control emphasized the positive effect of priming, as seen in T2 (thiourea @700 ppm) though it was variable in peroxidase activity and first year values of germination percentage (Table 1). The observed values on germination percentage were transformed into arcsine value for statistical calculation. In percent of germination, T6 (KNO3@0.2 Mpa) was highest only for 1st year and the peroxidase activity showed a positive response when treated with T3 (50 ppm gibberellic acid) followed by T11 (distilled water), T5 (0.1 Mpa KNO3) and T2 (750 ppm thiourea) under both year with pooled analysis. The anti-oxidative nature of peroxidase may favour the physiological aspect of seed, predominantly in germination. Normally, the role of bio-regulators is known for augmenting seedling expansion, mounting photosynthetic efficiency (Ramaswamy et al., 2007) etc. for emerged seedling.

Table 1: Seed priming influence on different parameters under physiological performances of seed.

Analysis of variance showed that all parameters were significantly influenced by seed priming for all the genotypes where speed of germination, vigour index and the enzyme, alpha amylase were predominant. The highest values were seen in V3 (RCM-FB-62) genotype for vigour index, alpha-amylase and non-significant deviation with the highest for seedling dry weight considering both years and pooled analysis (Table 2). In germination percentage, the V2 (Phyrngop dwarf) genotype showed highest for 1st year but, V(RCM-FB-62) indicated its dominance for 2nd year and pooled assessment (Table 2). The priming treatment might have an effect on peroxidase activity where V3 recorded lowest value as compared to other genotypes as specific genotypic nature was the most dominant factor. Considering the findings on speed of germination and seedling length, the genotype V1 exposed greater values among various genotypes. The important parameters vigour index and alpha-amylase under physiological performances of seed showed greater performance in V3 genotype, though it was not truly followed in seedling dry weight. In peroxidase activity, the observation was contrary to previous two parameters. Hence, these parameters may be considered for genotype evaluation.

Table 2: Deviation in genotypes considering physiological performances of seed and interactions of considerable two factors.

In interaction of treatment and genotype, the significant enhancement was noticed for parameters only in enzymatic actions considering both years and pooled analysis, but the non-significant trend was followed for most of the seedling parameters in full or partial manner except in seedling length (Table 2). Considering all parameters, the observed interacted values showed a general trend for upgradation in interaction of various priming over control (Lamichaney et al., 2018) irrespective of genotypes.
In Fig 1, the effect of treatment was visibly progressive for scheduled parameters considering their percent of enhancement over the control (T12). But, the activity of peroxidase demonstrated variable nature, highlighting highest effect in T3 with intermediate action in T2, T6 and T11. The most promising effect was observed in speed of germination, seedling dry weight and alpha amylase activity.

Fig 1: Deviance (in %) of various physiological performances of seed in priming over control.

The effect of primed seeds exposed a pronounced demarcation over control for most of the traits in the current study. It was reported that priming modified the metabolic processes involved in germination and earlier establishment of seedlings (Ghassemi-Golezani et al., 2008) as well as plants which may elicit the early development in comparison to control (Emongar, 2007). Moreover, the enzymatic action at initiation of germination may also be beneficial to advance seed vigour through sharing its expanding dry weight and length of the seedling (Arun et al., 2017).
In correlation matrix of the different physiological performances of seed, the R2 value of 0.7352 (Table 3) denoted the usual significant positive relationship for considerable characters. The positive significant mode was observed for most of the characters though the exception was revealed in peroxidase activity. The non-significant relationship was also followed in per cent of germination with seedling length, seedling dry weight and alpha-amylase activity though other parameters were highly significant within them in positive mode. All values of negative correlation showed non-significant differentiation particularly in case of peroxidase. However, peroxidase activity only showed a positive correlation with germination percentage, similar to a study by El-Araby et al., (2004). Therefore, the activity of peroxidase may favour germination by reducing antagonistic effect in seed, while alpha-amylase activity is closely related to germination activity. The adaptation of seed quality through priming and its effect on seedling parameters was studied by different scientists (Abnavi et al., 2012; Lamichaney et al., 2018). The dissimilar physiological performances of seed may diminish or increase in various ways through utilization of diverse treatments on seed in different crops, particularly under legumes and cereals. But seedling dry weight always showed an increase on application of PEG 6000 (Ghobadi et al., 2012), thiourea (Khan and Shah, 2010) KNO3 (href="#kumar_2018">Kumar et al., 2018), GA3 (href="#abnavi_2012">Abnavi et al., 2012; Arun et al., 2017; Mahnaz and Masomeh, 2016) similar to our study and sometimes in presence of CaCl2 (Afrayeem et al., 2018) and water as hydropriming (Mahnaz and Masomeh, 2016). Thiourea was the most capable sulphydryl group (-SH) of bio-regulator helpful in seedling growth (Anese et al., 2011 and Premaradhya et al., 2018). The intensity in activity of super-oxide dismutase (SOD) and quantity of soluble protein was amplified in radish owing to the effect of thiourea (LiQin et al., 2001). Growth promoters and nutrients directly influence metabolic activities in leaves during plant development and are thus responsible for improving the quality of the seed at the time of seed development. The potency of priming may enhance the activity of peroxidase, catalase, SOD and minimize electrolyte leakage under soaking.

Table 3: Correlation Matrix of the parameters allied to physiological performances of seed.

Generally, germination and interrelated parameters involve the activities of enzymes/isozymes, where anti-oxidative activity of specific isozymes is closely related to germination. A few reports relating to the valuable priming effects on seed indicated a build-up of more metabolites or osmotic adjustments at germination through the restoration of metabolic processes (Elkoca et al., 2007), or upgraded membrane integrity and superior physiological activities at germination (Sung and Chang, 1993). Soaked seed through hydro-priming or other modes of priming can supplement biochemical action like soluble protein, lipid, sugars, vitamin E, pro vitamin A etc. to bear the fundamental capability responsible for germination (Janeczko et al., 2015). Estimation of peroxidase activity can be used as an indicator of seed quality as reports have shown a sharp decline during aging (Pallavi et al., 2003). Seed treatment with thiourea has also been reported to enhance germination (Wahid et al., 2017) as observed in the present study.
The present interpretations showed considerable improvements in physiological performances of seed in French bean genotypes under the influence of priming. The treatment T2 (thiourea @ 750 ppm) exposed its promising potentiality in seedling irrespective of any genotype. V(RCM-FB-62) can be utilized to explore potentiality in cultivation predominantly in seed production programme to facilitate the quality seed of French bean.
The author would like to express her thanks to ICAR NEH Region, Umiam, Meghalaya for providing the original seed material used in the study.

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