Legume Research

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Legume Research, volume 44 issue 4 (april 2021) : 419-424

Seed germination response of commonbean (Phaseolus vulgaris L.) genotypes to optimal and sub-optimal temperatures

Amrit Lamichaney, T. Basavaraja, P.K. Katiyar, N.P. Singh
1ICAR-Indian Institute of Pulses Research, Kanpur-208 024, Uttar Pradesh, India.
  • Submitted22-01-2019|

  • Accepted08-04-2019|

  • First Online 14-08-2019|

  • doi 10.18805/LR-4122

Cite article:- Lamichaney Amrit, Basavaraja T., Katiyar P.K., Singh N.P. (2019). Seed germination response of commonbean (Phaseolus vulgaris L.) genotypes to optimal and sub-optimal temperatures . Legume Research. 44(4): 419-424. doi: 10.18805/LR-4122.
Rajmash is a cool season legume crop grown during rabi season in northern India. This crop is highly sensitive to low temperature stress during seed germination and seedling establishment stage. Therefore, identification of cold tolerant genotypes is essential in rajmash breeding programme. In the present study, 37 rajmash genotypes were subjected to germination at optimal (20oC) and sub optimal (10 and 15oC) temperatures in an incubator to understand the level of variation in germination and related at low temperature. At 20oC, the 37 genotypes recorded an average value of 98.47% with a minimum value of 91.35 %; while, at 15oC an average value of 96.02% and a minimum value of 86.10% and 10oC recorded an average value of 77.13% and a minimum value of 0%. This indicated that the genotypes could tolerate cold was recorded stress upto 15oC to a considerable level while, germination was hampered greatly at 10oC. Genotypes, EC150250, EC14920 and EC14351 showed the least reduction in germination and related parameters at sub optimal temperatures, hence these were identified to be cold tolerant at seed germination stage, which however, need to be validated under field condition.
Common bean/rajmash (Phaseolus vulgaris L.) is an important food grain legume, particularly in Latin America and in East and Southern Africa (Fetahu et al., 2014). In terms of nutrition, beans are often called the “poor man’s meat” for providing inexpensive source of a quality protein, essential minerals (iron and zinc) and vitamins (Beebe et al., 2000). In India, it is popularly known as rajma or rajmash, mainly grown by small and marginal farmers in the traditional production system (Rana et al., 2012; Sharma et al., 2012). Major rajmash growing Hilly states are Jammu and Kashmir, Himachal Pradesh, Uttarakhand and Sikkim where it is grown as kharif crop, it is also grown in plains of Uttar Pradesh, Bihar, Maharashtra, Tamil Nadu, Kerala and Karnataka during rabi season (Kumar et al., 2009). Since, rajmash is primarily grown during rabi season in northern plain region, it exposes seed germination and seedling establishment stage to low temperature. Rajma is extremely sensitive to mean temperatures below 15°C at seed germination and seedling development stage (Vandana et al., 2015; Basavaraja et al., 2017). Prevalence of low temperature stress at seeding (late sown crop) lead to delayed emergence and slow vegetative growth resulting in poor biomass production and ultimately causing reduction in grain yields (Antonio et al., 2016).
 
In past years, breeding objectives was focused on developing disease resistant high yielding varieties with improved plant architecture and seed quality. However, little efforts were given for development of cold tolerant cultivar. Nevertheless, few reports are available regarding genotypic variation for cold tolerance in common bean. Dickson (1971) observed that white seeded rajmash were more susceptible to cold stress during germination as compared to coloured genotypes. Seed colour is known to be associated with differences in seed vigour especially in legumes, whereby, coloured seeds are reported to have better seed vigour (Lamichaney et al., 2016; Lamichaney et al., 2017). Balasubramanian et al., (2004) reported genotypic differences in cold tolerance among the wild and cultivated rajmash. Temperature is an important limiting factor for rajmash production as low temperature during sowing can not only reduce emergence but could delay substantially the germination and seedling emergence. Therefore, development of varieties tolerant to cold stress especially during germination and seedling emergence is the need of the hour to increase the adaptability of rajmash to other non-traditional areas of the country; for which screening and identification of genotypes tolerant to sub-optimal temperatures is a must. Since, rapid and uniform seed germination is the best selective indicator of cold tolerance, present investigation was conducted to screen rajmash genotypes for their ability to germinate at optimal (20°C) and sub-optimal low (10° and 15°C) temperature.
Plant material
 
A total of 37 rajmash genotypes (5 released varieties and 32 germplasm accessions) were investigated for cold tolerance during germination stage. Seeds used were harvested in the month of April 2016 and were stored at ambient temperature in air tight seed storage bin. A pure, undamaged and uniform sized seed from each genotype was selected for cold tolerance study. The detailed information of 37 genotypes with respect to growth habit, other seed traits such as seed colour, shape, hilum colour, length, width and 100-seed weight are given in Table 1.
 

Table 1: Category, growth habit, seed colour, seed shape, hilum colour, seed length, seed width and 100-seed weight of rajmash (French bean) genotypes studied.


 
Screening for cold tolerance
 
For cold tolerance evaluation, seeds of 37 rajmash genotypes were germinated under three temperatures: 10, 15 and 20°C, 20°C being considered optimum temperature for germination of rajmash (Anonymous, 2015). Twenty-five seeds of each genotype were placed in 13 cm Petri-plates lined with pre wetted filter paper. Seeds were surface sterilized with 0.5 % sodium hypochloride followed by rinsing with deionized or distilled water for 2 to 3 times. The Petri-plates were transferred to germinator maintained at 10, 15 and 20°C. The experiment was conducted in a complete randomized block design with four replicates. Seedling germination was counted daily. The seeds were considered germinated when radicle attained approximate length of 2 mm. At the end of the experiment, the final percentage germination and actual time to 1% germination or time taken to initiate germination (T1), actual time to 50% germination (T50), uniformity of germination or time between 25 and 75% germination (U7525) and mean germination time were calculated. The daily germination performance of rajmash seeds at different temperatures were interpreted by “Germinator curve-fitting1.29.xls” microsoft excel model” (Joosen et al., 2010) using the following equation (El-Kassaby et al., 2008).
 
 
Where,
Y is cumulative germination percentage at time X, Y0 is intercept on the Y axis, a is maximum germination per cent, b is steepness of the curve and c is time taken by 50% of seeds to germinate (T50). The arcsine transformed germination data, T1, T50, U7525 and MGT data were statistically analysed by completely randomized one-way ANOVA using online software OP-STAT (Sheoran et al., 1988).
Germination is influenced by many factors, of which temperature, relative humidity and moisture are the key determinants that drives the process of germination. Each crop has its own temperature requirement or optimal temperature that facilitates rapid and uniform germination. However, increase or decrease in temperature form optimal could drastically affect not only final germination per cent but also may increase the time taken to achieve germination (Rodino et al., 2007). Also time taken from first germination to final germination (uniformity of germination) of a seed lot may increase with fluctuation in optimal temperature which results into non uniform plant growth making inter-cultural operation difficult. For successful and uniform germination of rajmash 20°C is considered optimum (Anonymous, 2015). Therefore, two sub-optimal temperatures i.e. 10° and 15°C was included in the present investigation to understand the level of cold tolerance in a set of 37 rajmash genotypes. At optimal germination temperature (20°C), there were no significant differences in the final germination and time taken to achieve first germination (T1) among 37 genotypes, however, these genotypes differed significantly in time taken to achieve 50% germination (T50), uniformity of germination (U7525) and mean germination time (MGT) (Table 2). At optimal temperature (20°C), all the genotypes recorded germination percent of >91%, minimum recoded in genotype IC 311670 and maximum (100%) in 12 genotypes (GPR 203, EC 400445, ET 8405, PDR 14, ET 8447, BSLR 2, NO 3160A, BLF 101, UTKARSH, EC 400414, ET 8409 and EC 400401). Time to achieve first germination in 37 genotypes at 20oC varied between 32.11 to 101.38 h with a mean of 67.87 h. Likewise, time taken to achieve 50% germination varied between 67.44 (EC 500232) to 206.26 h (EC 400361) with a mean of 104.58h. Similarly, uniformity of germination (U7525) and MGT at 20°C among 37 genotypes ranged between 2.92-79.93 h and 68.15-208.09, respectively (Table 2 and Table 3).
 

Table 2: Mean germination per cent, time taken to initiate germination (T1), time taken to achieve 50% germination (T50), time between 25 and 75% germination (U7525) and man germination time of 37 genotypes at optimal (20oC) and sub-optimal (10 and 15oC) temperatures.


 

Table 3: Variability in germination and related parameters of 37 genotypes at optimal (20oC) and sub-optimal (10 and 15oC) temperatures.


 
At first sub-optimal temperature (15°C), genotypes recorded mean germination percent of 96%, minimum (86.10%) recoded in genotype EC 564795 and maximum (100%) in 11 genotypes (ET 8404, ET 8456, BD 9116291, IC 14351, BSLR 2, BLF 101, UTKARSH, EC 14920, EC 400414, EC 150250 and EC 400401). Time to achieve first germination in 37 genotypes at 15°C varied between 50.61 to 133.94 h with a mean of 92.13 h. Likewise, time taken to achieve 50% germination varied between 105.42 (EC 150250) to 172.39 h (IC 25537) with a mean of 137.35h. Similarly, uniformity of germination (U7525) and MGT at 15°C among 37 genotypes ranged between 9.06-63.94 h and 108.06-171.31, respectively (Table 2 and Table 3).
 
At second sub-optimal temperatures (10oC), two genotypes (ET 8447 and AMBER) recorded 0% germination (100 % reduction from 15 and 20°C) and one genotype (EC 400414) recorded 10% germination (90 % reduction from 15 and 20°C). The germination per cent at 10°C varied between 0-100% with a mean of 77.13%. Genotypes ET 8447 and Amber did not germinate therefore they do not have value for T1, T50, U7525 and MGT. Likewise, there are few genotypes which germinated but did not achieve 50% germination, so they have T1 value but not T50. Time to achieve first germination in 35 genotypes at 10°C varied between 99.02 to 425.22 h with a mean of 181.23 h. Likewise, time taken to achieve 50% germination in 31 genotypes varied between 173.18 to 398.18 h with a mean of 272.55 h. Similarly, uniformity of germination (U7525) and MGT at 10°C among 35 genotypes ranged between 12.95-131.55 h and 114.32-382.38, respectively (Table 2 and Table 3).

The proportion of germination at optimal temperature (20°C) had an average value of 98.47% and a minimum value of 91.35 % while first sub-optimal temperature (15°C) had an average value of 96.02% and a minimum value of 86.10% and second sub-optimal temperature (10°C) it had an average value of 77.13% and a minimum value of 0%, which indicates that the genotypes could tolerate cold stress upto 15°C to a considerable level while germination was hampered greatly at 10°C. The emergence was delayed at sub-optimal temperatures by 137.02 and 35.74% at 10 and 15°C, respectively as compared to optimal temperature (Table 2). Genotypes, ET 8447 and Amber were the most susceptible to cold stress as they failed to germinate at 10°C, which otherwise recorded 94.8 and 97.3 % respectively at 15°C and 100 and 96% respectively at 20°C. Likewise, time to achieve 50% germination was delayed by 160.61 and 31.33 times at 10 and 15°C as compared to 20°C, respectively (Fig 1). At 20°C, time taken between 25 and 75% germination to occur (uniformity of germination) was 21.80 h which was increased by 23.39% times at 15°C and by 119.7% times at 10°C (Table 2). Our findings corroborated with Scully and Waines (1987) who reported that rajmash genotypes took about 16-24 days to emerge at sub optimal temperature of 12°C. It has been reported that the activity of alpha amylase gets reduced at low temperature (Du et al., 2010). The reduction in germination value and increase in the time taken to germinate or attain 50% germination at sub optimal temperature in rajmash may be due to decrease in alpha amylase activity which is essential for breaking down starch and provide food material for germination to occur. At higher temperature the germination rate, speed of germination and seedling emergence will be higher and faster which have been reported in different crops including food legumes, such as soybean, cowpea, groundnut and chickpea (Craufurd et al., 1996; AwalandIkeda, 2002; Lamichaney and Katiyar, 2017). Slow germination and emergence of rajmash seed at low temperature might attract fungal infection which ultimately leads to seed death before emergence causing poor field emergence and low productivity. Delayed emergence of seeds due to low temperature may lead to damping off diseases which are caused by several soil and seed borne fungi like Pythium, Botrytis, Rhizoctonia, Cylindrocladium, Fusarium, Phoma, Alternaria, Phytophthora etc (Rodino et al., 2007).
 

Fig 1: Effect of temperature on rajmash seed germination and seedling growth.

The results indicated that the rajmash genotypes could tolerate cold stress upto 15°C to a considerable level in terms of germination and related parameters while, germination and related germination parameters was greatly affected/ delayed at 10°C. The promising cold tolerant genotypes (EC150250, EC14920 and EC14351) identified could be used in rajmash breeding programme.

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