Legume Research

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Research Article

Legume Research, volume 43 issue 3 (june 2020) : 415-420

Effect of Boron Priming on the Seed Vigour in Different Varieties of Alfalfa (Medicago sativa L.)

F.S. Xia1, Y.C. Wang1, F. Wang1, C.C. Wang1, H.S. Zhu1, M. Liu1, Y.M. Huai1, K.H. Dong1,*
1College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province 030801, China.

  • Submitted08-03-2019|

  • Accepted12-05-2020|

  • First Online 25-06-2020|

  • doi 10.18805/LR-484

Cite article:- Xia F.S., Wang Y.C., Wang F., Wang C.C., Zhu H.S., Liu M., Huai Y.M., Dong K.H. (2020). Effect of Boron Priming on the Seed Vigour in Different Varieties of Alfalfa (Medicago sativa L.) . Legume Research. 43(3): 415-420. doi: 10.18805/LR-484.

ABSTRACT

This experiment was designed to determine the influence of boron priming on seed vigour in thirteen varieties of alfalfa (Medicago sativa L.). Alfalfa seeds were primed with 1.8% (W/V) concentration of borax solution for 0, 3, 6, 9, 12 and 24 h at 20°C. The results showed that the original vigour of alfalfa seeds was disparate in thirteen varieties and there were highly significant (P<0.01) differences observed in both varieties and priming time and their interactions on the vigour of the seeds. Alfalfa seeds of WL525HQ and WL656HQ might be more sensitive to boron toxicity, but seeds of WL298HQ, WL343HQ, WL354HQ, WL903 and Pianguan were insensitive to boron toxicity. Thus, it is necessary to carefully select appropriate varieties of alfalfa seeds for the application of boron priming.

INTRODUCTION

Alfalfa (Medicago sativa L.) has more excellent features comparing with other forage crops, e.g. high yield, rapid regeneration, high adaptability, rich nutrition and low production cost (Zhang et al., 2019). Therefore, it is considered the most important forage crop and is extensively cultivated primarily for hay, silage and pasture in animal feeding around the world (Hawkins and Yu, 2018; Yang et al., 2019). Furthermore, alfalfa can also be used to improve soil fertility as an important green manure crops (Zhang et al., 2017a). Currently, the cultivation of alfalfa is increasing rapidly in the world, especially in arid and semi-arid areas (Huang et al., 2018). Hence, the quantity and quality of alfalfa seeds should be improved correspondingly. However, the amount of alfalfa seed fields is not growing at a sufficient rate and the yield of alfalfa seeds is still relatively low (Zhang et al., 2017b), this seriously restricts the splendid development of alfalfa industry around the world. Consequently, how to improve alfalfa seed yield has become a world problem to promote the rapid development of alfalfa industry.
        
Boron is an important micronutrient for vascular plants. In recent years, it has been used more widely in order to increase grain yield in crop production (Parry et al., 2016; Iqbal et al., 2017). Likewise, alfalfa seed yield can be effectively improved using foliar application of boron (Du et al., 2009). Nevertheless, there are still few studies on the application of boron to improve alfalfa seed yield and it is especially scarce for research on the optimal application methods. Compared with soil application and foliar spray, seed priming has been emerged as an attractive and easy physiological strategy in micronutrients application (Farooq et al., 2012; Arun et al., 2017). Currently, it is necessary to discover and optimize new priming agents in order to satisfy the cheaper, exercisable and easily accessible requirements of farmers (Hussain et al., 2018). Seed priming with boron has been shown to be effective in encouraging seedling emergence, growth and grain yield during crop production (Iqbal et al., 2017). Moreover, our previous study also found that appropriate priming with boron can improve the seed vigour of alfalfa (Xia et al., 2019). Nonetheless, the understanding remains poor in response of boron priming on seed vigour of different varieties of alfalfa.
        
Thus, the objectives of this research are to compare the changes in germination percentage, germination index, mean germination time and seedling vigour index in different varieties of alfalfa after boron priming, to evaluate the response of boron priming on seed vigour in different varieties of alfalfa and to accurately provide an exercisable, effective and economic method for the application of boron in alfalfa seeds’ production.

MATERIALS AND METHODS

Seed source
 
Thirteen varieties of alfalfa seeds were used in this experiment, including Pianguan, WL168HQ, WL298HQ, WL319HQ, WL343HQ, WL354HQ, WL363HQ, WL366HQ, WL440HQ, WL525HQ, WL656HQ, WL712 and WL903. Alfalfa (variety: Pianguan) seeds were collected by the Forage Seed Laboratory of Shanxi Agricultural University in August 2017 and then sealed in plastic bags and stored at -20°C. Other varieties were provided by Beijing Rytway Ecotechnology Co., LTD. The experiment was launched on July 2018 in the Forage Seed Laboratory of Shanxi Agricultural University.
 
Boron priming treatments
 
Alfalfa seeds were soaked in 1.8% (W/V, selected according to 0, 0.1, 0.3, 0.6, 1.2, 2.4 and 4.8%) concentration of borax solution for 0, 3, 6, 12 and 24 h at 20°C. Thereafter, seeds were immediately scoured two times with deionized water and surface moisture were removed using filter paper. They were then air-dried for three days in the dark at 25°C and 45% RH in order to ensure moisture content reached approximately 10% (on fresh weight basis). Each treatment had four replicates.
 
Germination tests
 
Germination tests were executed following the ISTA rules (2017). Hundred seeds from each sample were selected and placed into petri dishes that was lined with three filter papers and wetted with 10 ml deionized water. They were then placed in constant temperature incubators at 20°C. Germination was recorded daily based on 2 mm radicle growth through seed coat. The number and length of normal seedlings were counted in each petri dish on day-10. Germination percentage was computed by the method of ISTA (2017). Germination index and seedling vigour index were calculated according to Abdul-Baki and Anderson (1973). Mean germination time was counted according to Ellis et al., (1982). The full formula had been reported in our previous studies (Xia et al., 2019).
 
Statistical analyses
 
Seed germination data was respectively tested using the Kolmogorov-Smirnov test and the homogeneity test of variances and was conformed to normal distribution and homoscedasticity. Mean difference of comparisons were performed using an analysis of variance (ANOVA), which was conducted using SPSS for Windows ver. 13.0 followed by Duncan’s multiple range test (P = 0.05).

RESULTS AND DISCUSSION

Effects of boron priming on germination percentage
 
The initial germination percentage of alfalfa seeds was different across varieties and their variations were also different with the extension of boron priming times (Table 1). There were highly significant (P<0.01) differences among varieties, priming time and their interaction on germination percentage of alfalfa seeds (Table 2). In this study, germination percentage of WL525HQ and WL656HQ declined significantly (P<0.05) with increasing length of priming time, which manifested that priming with 1.8% boron caused a decrease in the germination percentage of alfalfa seeds in both varieties. Excessive boron could inhibit both the vegetative and reproductive growth of plants (Kaya and Ashraf, 2015). Also, our previous reports showed that the germination percentage of alfalfa seeds could be markedly reduced with immoderate boron priming (Xia et al., 2019). Therefore, these results showed that the seeds of WL525HQ and WL656HQ might be sensitive to boron toxicity in terms of germination percentage. Nonetheless, the germination percentage of WL366HQ and Pianguan reached the maximum level at 3 h, which was significantly (P<0.05) higher than those unprimed seeds, this proved that priming with 1.8% boron promoted the germination percentage of alfalfa seeds in both varieties. Li et al., (2017) found that seed priming could improve the germinating ability of alfalfa seeds. Similarly, previous studies also indicated that boron priming could effectually promote seed germination (Deb et al., 2010; Iqbal et al., 2017). Beyond this, a significant (P<0.05) decrease occurred in those seeds primed for 12 and 24 h. These results indicated that the seeds of WL366HQ and Pianguan might be insensitive to boron toxicity in terms of germination percentage. This was similar to our previous reports in those seeds primed with 1.2 and 2.4% boron (Xia et al., 2019). Likewise, strong boron priming could not promote the seed germination (Farooq et al., 2012). In this study, the difference in germination percentage was not significant (P>0.05) between those primed for 0 and 3 h in other varieties of alfalfa seeds. Particularly, the germination percentage of WL319HQ, WL343HQ and WL354HQ still maintained a high level after being primed at 0, 3 or 6 h and there were no significant (P>0.05) differences among these seeds. These results showed that these varieties were moderately sensitive to boron toxicity in terms of germination percentage.
 

Table 1: Effect of boron priming on germination percentage in different varieties of alfalfa seeds.


 

Table 2: Variance analysis of priming times and varieties on alfalfa seed vigour.


 
Effects of boron priming on germination index
 
The original germination index of alfalfa seeds was disparate in different varieties and their changes were also varies with the extension of boron priming times (Table 3). There were highly significant (P<0.01) differences among varieties, priming time and their interaction on germination index of alfalfa seeds (Table 2). Seed priming had been regarded as an effective method to improve germination index of seeds (Xia et al., 2017). In this study, the germination index of WL903 and Pianguan reached the maximum level at 3 h and they were significantly (P<0.05) higher than others. These results indicated that the germination index of alfalfa seeds in both varieties were promoted by priming with 1.8% boron. This might be attributed to the complement of antioxidant systems and ultrastructure of embryonic cells during seed priming (Xia et al., 2017). Moreover, a significant (P<0.05) decrease occurred in these two varieties of alfalfa seeds primed for 6 to 24 h. Therefore, these results showed that these two varieties of alfalfa seeds were insensitive to boron toxicity as far as germination index. Nonetheless, the germination index reduced markedly (P<0.05) with the extension of priming time in other varieties of alfalfa seeds, which showed that these seeds might be more sensitive to boron toxicity according to their germination index.
 

Table 3: Effect of boron priming on germination index in different varieties of alfalfa seeds.


 
Effects of boron priming on mean germination time
 
Seed priming has been deemed to activate hydrolytic enzymes and improve embryonic physiology, thereby their germination happen in less time (Bam et al., 2006). Additionally, boron could promote the remobilization of their stored nutrients during seed germination (Bonilla et al., 2004). In this study, there were highly significant (P<0.01) differences among varieties, priming time and their interaction on mean germination time of alfalfa seeds (Table 2). The mean germination time of WL903 reached the minimum level at 3 h, which was significantly (P<0.05) lower than others (Table 4). These results showed that the germination speed of WL903 was accelerated by priming 3 h with 1.8% boron. Nevertheless, they decreased significantly (P<0.05) after priming from 6 to 24 h. Moreover, mean germination time declined prominently (P<0.05) with the extension of priming time in other varieties of alfalfa. Seed priming was based on the relationship between imbibition and water potential (Bewley et al., 2013). However, the increase in concentration of priming solution usually leads to a decline in water potential delaying cell activation (Xia et al., 2017). Thus, these results illustrated that these varieties of alfalfa seeds might be more sensitive to boron toxicity in terms of mean germination time. This was similar to our previous reports (Xia et al., 2019). Farooq et al., (2011) also found that boron priming could distinctly delay the emergence of rice seeds.
 

Table 4: Effect of boron priming on mean germination time indifferent varieties of alfalfa seeds (days).


 
Effects of boron priming on seedling vigour index
 
The level of seedling vigour index generally demonstrated the ability of seedling establishment in growth process. Iqbal et al., (2017) found that boron priming at low concentration resulted in seedlings with longer shoots and consequently able to acquire more nutrients. However, there were highly significant (P<0.01) differences among varieties, priming time and their interaction on seedling vigour index of alfalfa seeds (Table 2). In this study, seedling vigour index increased in the varieties of WL168HQ, WL298HQ, WL319HQ,WL343HQ, WL363HQ, WL366HQ, WL712 and Pianguan after 3 h of priming, which was significantly (P<0.05) higher than those of non-primed (Table 5). These results showed that seedling vigour index of these varieties were accelerated by priming 3 h with 1.8% boron. Particularly, seedling vigour index of WL298HQ, WL343HQ and WL354HQ remained at the highest level after 3 h of priming, showing that these three varieties of alfalfa seeds were insensitive to boron toxicity. The successful establishment of a seedling relied largely on effective mobilization of storage nutrient (Sew et al., 2016). Hence, primed seeds might possess greater energy to complete seedling growth (Chen and Arora, 2013). Nevertheless, they all declined with the extension of time, which indicated that excessive boron priming might also inhibit the growth of alfalfa seedlings. Our previous study had similar results (Xia et al., 2019). Unlike these varieties, seedling vigour index of WL440HQ, WL525HQ, WL656HQ and WL903 declined with increased priming time, which illustrated that seedling vigour index in these varieties of alfalfa seeds might be more sensitive to boron toxicity.
 

Table 5: Effect of boron priming on seedlings vigour index in different varieties of alfalfa seeds.

CONCLUSION

There were highly significant (P<0.01) differences among varieties, priming time and their internation on germination percentage, germination index, mean germination time and seedling vigour index of alfalfa seeds. Additionally, the seeds of the varieties viz., WL525HQ and WL656HQ were more sensitive to boron toxicity, but the varieties viz., WL298HQ, WL343HQ, WL354HQ, WL903 and Pianguan were insensitive.

ACKNOWLEDGEMENT

This research was financially supported by the Natural Science Foundation of China (NSFC31702171), the key research and development program of Shanxi (201903D221091) and Science and Technology Innovation Fund of Shanxi Agricultural University (2016YJ15).

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