EFFECTS OF PROBIOTIC AND ORGANIC MANURES FOR SOIL AMENDMENTS ON SEEDLING SURVIVABILITY AND LEAF CHLOROPHYLL FOR CROP IMPROVEMENT IN WHEAT (TRITICUM AESTIVUM L.)

Under this study a total of nine treatments were considered for soil amendments where organic manures, chemical fertilizer and probiotics were used to evaluate the efficiency on growth and yield of wheat. On the basis of seedling disease incidence and chlorophyll presence parameters data were recorded on seedling infection at 14 days, seedling blight at 21 days, and chlorophyll content at 70 days after sowing. In this study the same experiments were conducted up to three years (2016-2019) and data were compiled and analyzed. It was observed that the disease prevalence attributes significantly decreased in T5 (Trichoderma harzianum + vermicompost + green manuring) than other treatment combinations and control (T0). Noteworthy results were found on seedling health, application of probiotic and organic manures which significantly influenced the less infection of wheat. Besides this, chlorophyll content of wheat leaf and plant number with effective tiller enhanced largely under T8 (chemical fertilizer) than T4 (poultry manure + vermicompost + green manuring) and control (T0). The main advantage of this research is to evaluate efficiency for growing healthy wheat plant which reflects on plant growth and yield and ultimately helps for food security.


Introduction
Wheat (Triticum aestivum L.) is a staple food of billions of people all over the world and third most agricultural commodity next to rice and maize. The contribution of wheat is being increased significantly in different countries as Nigeria (1% -6.64%), India (11.85% -20.41%) and China (12.20% -17.83%) regarding total kcal (Shewry and Hey 2015). In Bangladesh, wheat is one of the important grain crops regarding economic and consumption purposes. Wheat is used as industrial and commercial products for human consumption having high protein with others nutrients that is also a cheaper source of feed for livestock (Hammad et al. 2011). Land engaged for cultivation decreased with time to meet up the accommodation demand for geometric growth of population. To face the land limitation problem and provide food for increased population, sustainable use of land is burning issue for agricultural development (Haque andIslam 2018, Alam et al. 2020). The substantial pressure on land may create various constraints due to imbalanced and inadequate application of fertilizer (Kumar and Abraham 2018). Chemical fertilizer may increase the wheat yield but prolonged application resulted human health hazard and imbalanced natural resources (Hossain et al. 2021a). So, need to link soil management and cropping design that are more environmentally sustainable (Karnel et al. 1994). Though transition period of three years produce lower performance but higher soil fertility found over time in conventional to organic farming (Tester 1990). So, for achieving higher yield and retain soil health sound organic amendments are felt to be important. A good soil should have organic matter content of more than 2.5%. Most of the soils in Bangladesh have low to very low organic matter content, generally less than 1.5% (BARC 2012). Soil productivity is greatly influenced by the application organic manures like crop residues, green manure and animal manure in the soil-plant system (Yadvinder-Singh et al. 2008, Hossain et al. 2021a. Probiotics are live microorganisms which confer a beneficial health benefit on the host. Trichoderma is considered as a probiotic having the characteristic of fungi and considered as a potential and promising bio-control agent and growth promoter for many crops. Bio-fertilizer is regarded as an eco-friendly substitute of chemical fertilizer trends to enhance soil fertility and increase crop productivity and yield without creating harm to the environment (Hermosa et al. 2012, Hossain et al. 2021b. It was established that Trichoderma is a biological degrader and promotes plant disease defense, increasing the immunity of the plant. Hence, Trichoderma have tremendous opportunities for disease management of soil borne pathogens but also have the capacity to improve plant growth parameters and soil health. It is authentic that fairly good soil fertility and plant nutrients are important to farming. Hence to improve soil fertility and increase plant nutrients availability, efforts need to be made to increase soil organic matter content. So, in view of the above realities an initiative was undertaken to focus the probiotic and organic manure performance on wheat seedling and plant growth resulted on yield, which may help to increase the productivity of wheat.

Soil and field preparation
The present research work has been carried out in the Experimental Field of the Institute of Biological Sciences (IBSc) and laboratory works has been carried out at Plant Biotechnology and Genetic Engineering Lab., IBSc and Plant Pathology Lab., Department of Agronomy and Agricultural Extension, University of Rajshahi, Bangladesh during the period from July 2016 to June 2019. The experimental field geographically situated at 24°17` N latitude and 88°28` E longitude at an elevation of 20 m above the sea level belonging to the Agro-Ecological Zone-11 (AEZ-11). The soil of the experimental field has characteristics like poorly drained with moderately permeability, loam and slightly alkaline (pH = 8.10) in nature. The physical properties of the experimental soil had moisture content 19.40%, particle density 2.65 g/cc, bulk density 1.27 g/cc and porosity 51.34%. Furthermore, chemical properties like the organic matter content 1.20%, K 0.150 cmol (+)/kg, total N 0.07%, P 26.30 µg/g, S 12.50 µg/g, Zn 0.75 µg/g, EC 145 µs/cm and C:N 10:1 were recorded.

Seedling infection
The seedlings which were found yellow and rotted at the base are considered as infected. From germination up to 14 days regular observation was made and infected seedlings were counted in each plot. Infection was estimated by the following formula:

Seedling blight
Seedlings which were dead and became straw in color defined as seedling blight. Seedling infection is treated as the first step of seedling blight. Thus, blighted seedlings were counted at 21 DAS in every plot. It was estimated as by the following formula:

Leaf chlorophyll content
The atLEAF handled chlorophyll meter was used to assess leaf greenness. The meter determines light transmittance through the leaf at 660 and 940 nm wavelengths. The readings were obtained by inserting the middle portion of the topmost fully expanded leaf in the slit of the meter. At least three readings from leaves of randomly selected plants in each plot were recorded and mean value was determined. Abnormally looking or insect attacked plants were not selected for measurement (Ali et al. 2020).

Total plant/m 2
Before harvesting 1 m 2 area was selected with quadrate and the number of total plants was counted. This operation was done in each plot preferably choosing the five rows in the middle of the plot.

Effective tiller/plant
The numbers of tillers bearing panicles were counted at harvest with the help of quadrate from 1 m × 1 m area, which expressed as effective tillers/plant.

Statistical analysis
The recorded data were compiled and tabulated for statistical analysis. The data were analyzed statistically for analysis of variance (ANOVA) and Duncan's Multiple Range Test (DMRT) through r-studio by the "Agricolae" package.

Seedling infection
Regarding various treatments effect, the lower rate of seedling infection (0.32%) was found after application Trichoderma harzianum (T5) in comparison with the maximum infection from T0 (2.32%) in 2018-2019. All the treatments in the experimental period presented significant differences among each other. But T1 (rice straw + vermicompost + green manuring) and T8 (chemical fertilizer) was statistically identical. Reduction of seedling infection at 14 DAS under different soil amendments showed more or less the following order as T5>T7>T4>T3>T2>T6>T1>T8>T0 during the whole research period shown in Table 1.

Seedling blight
Under this study, recorded results showed that there was a significant variation among the treatments. The highest inhibition effect against seedling blight (0.87%) was observed by treatment T5 and the lowest (4.74%) from control (T0) during 2018-2019 ( Fig.1 a-b). Next to treatment T5, the treatment T7 also showed remarkable effect in reducing seedling blight (1.07%) which was statistically identical (1.09%) with T4 (Table 1). Fig. 1(a-b): Seedling status and plant growth by adding probiotic and organic manures for soil amendments, a) seedling infection and, b) seedling blight.

Leaf chlorophyll content
A significant difference was observed from various soil amendments for the chlorophyll content of wheat leaf at 70 DAS. The highest value of chlorophyll content (51.48 SPAD) was recorded from chemical fertilizer (T8) and which was followed by T4 (48.37 SPAD) and T5 (48.08). In this regard, the smallest value (40.33 SPAD) was noticed from T0 during the season 2018-2019. Due to soil amendment with chemical fertilizer (T8) a b provided more chlorophyll content and the trend was maintained from beginning to the end of the study (Table 1). Each value represents the average of three replicates. In the column, mean values bearing similar letter(s) or without letter are identical and those having dissimilar letters are differed significantly as per Duncan's Multiple Range Test. T0 = control, T1 = rice straw + vermicompost + green manuring, T2 = cow dung + vermicompost + green manuring, T3 = compost + vermicompost + green manuring, T4 = poultry manure + vermicompost + green manuring, T5 = T. harzianum + vermicompost + green manuring, T6 = mung bean residues + vermicompost + green manuring, T7 = T. viride + vermicompost + green manuring, T8 = chemical fertilizer, SPAD = The soil plant analysis development. LS = level of significance, ** = 1% level of significance, CV = co-efficient of variation.

Total plant/m 2
Total plant/m 2 at harvest varied significantly due to different soil amendments all over the study period. The highest frequency of total plant/m 2 was found in T8 (199.11%) and the lowest result was recorded in T0 (158.67%) in 2016-2017 (Fig. 2). The next highest value was recorded from T4 (191.56%) and T5 (

Effective tiller/plant
Under this study, recorded results on effective tiller/plant showed significant difference by the influence of treatments. Out of different soil amendments, chemical fertilizer (T8) and poultry manure (T4) performed as superior to the rest of the treatments mainly unamend (T0) all over the research period (Fig. 3). On the basis of the above statement, T8 was produced the maximum number of effective tiller/plant (3.49%) followed by T4 (3.40%), whereas the minimum value (2.07%) was given by T0 in 2016-2017. During 2017-2018, treatments were exhibited approximately equal performance with slight increase of effective tiller/plant. However, 3.64% and 2.10% were the greater and less value of effective tiller/plant obtained by treatment T8 and T0. In the last year 2018-2019, the maximum value on this parameter was found from T8 (3.89%) and T4 (3.69%) and then gone downward as followed T5 (3.36%) >T7 (3.22) >T3 (3.09) >T2 (2.91%) >T6 (2.84%) >T1 (2.73%) over T0 (2.16%).

Seedling infection
A significant variation of seedling infection was noticed all over the research period. The trend of infection was found to decline with the progress of the study due to application of probiotic and organic manure. The highest rate of seedling infection was found in T0 (2.32%), while the minimum rate of infection was recorded from T5 (0.32%) in 2018-2019. It might be due to the definite role of probiotic fortified-organic manures that exerted antagonistic effect against the fungi causing seedling infection. Furthermore, the addition of organic matter may create a hospitable environment for growth and development of fungal antagonists resulting in enhanced biocontrol activity (Sarkar et al. 2002). This finding was in harmony with the reports Tewari and Rajbir (2005).

Seedling blight
Seedling infection later on converted into seedling blight. Like seedling infection, this parameter was also varied significantly under various treatments all over the study period. The control activity against seedling blight was highly observed by T5 (0.87%) and the lowest exhibited from check T0 (4.74%) during 2018-2019. Such result may be obtained for the efficacy of the prohibitory and biocontrol action of the treatment. This efficacy of Trichoderma fortified-organic manures was evaluated to reduce the pre-emergence and post-emergence seedling mortality, diseases of stem and root of chickpea by Talukder et al. (2017). Similar control activity of Trichoderma spp. against plant pathogen and seedling mortality was also reported by Bhuiyan et al. (2007) and Ha 2010.

Leaf chlorophyll content
Chlorophyll content of leaf from diverse treatments was provided significant dissimilarities from beginning to end of the study. On the focus of organic amendments, treatment T4 (48.37 SPAD) next to T8 (51.48 SPAD) had the bigger accumulation of chlorophyll and the smaller was noticed from T0 (40.33 SPAD). It may probably relate with the decomposition rate of organic manures that happened in case of T4 treatment. This result was supported by Khan et al. (2005) with the statement that N supply in organic treatment is generally restricted and slows N mineralization as compared to crop demand of N. Chlorophyll content is closely related to the presence of N and poultry manure (T4) had the higher ability of supplying N. This result is an agreement with the findings of Sims and Wolf (1992), Khanam et al. (2001), Ru et al. (2012) and Krishna (2013).

Total plant/m 2
The parameter plant/m 2 is one of the major vegetative growth indicators of wheat. The best values of plant/m 2 were obtained from T8 (216.11%) and then T4 (202.81%) where the lowest at T0 (158.66%).
The ability of such enhancement plant/m 2 due to application of poultry manure might contain high amounts of nutrients and organic matter that improved the soil structure and environment which aids density of plant. Result in the study was in the same manner previously notified by Hassan (2002), Dauda et al. (2008) and Ismaeil et al. (2012).

Effective tiller/plant
Tillering is one of the important growth stages of wheat which provides the necessary stalks for satisfactory production. Though tillers are lateral branches which emerge from the main stem, but not all produce an ear. Generally, the first two-three tillers are noted as productive tillers of wheat and production is related to environment and nutrition supply. In this investigation, a greater number of productive tiller/plant was demonstrated by T8 (3.89%) and T4 (3.69%). This advancement of effective tiller might be due to optimum moisture and available nutrients provided by poultry manure which may have created the favorable condition for wheat growth. This was supported by Simpson (1990), Eck and Stewart (1995), Mitchell and Tu (2005) and Sistani et al. (2008). The result of this investigation was related to the observations of Belefant-Miller (2007), Abbasi and Khaliq (2016).

Conclusion
Quick response of chemical fertilizer offered an instant results nevertheless it has harmful effect not only crops as well as soil health. Therefore, organic soil amendment has given emphasis in this study. Seedling survivability was found greater by the application of probitic combination, whereas potential and sustainable improvement of wheat growth with organic soil amendment was found. Regarding this findings, Trichoderma harzianum, green manure and vermicompost may be useful for against seedling mortality and in addition of green manure, vermicompost and poultry manure may be a worthy substitute for wheat growth. Data of this study revealed that the probiotic particularly Trichoderma harzianum produce healthier seedling and poultry manure exhibited the capacity of supply nutrient demand and chlorophyll synthesis for improvement in growth of wheat as compared to other manures.