INTRODUCTION
Tubtim Chum Phae rice
Tubtim Chum Phae rice, often referred to as the gem of Thai rice, is a new strain developed through the hybridization of Hom Mali rice and Sung Yod Patthalung rice Hom Mali rice is renowned for its exceptional flavor, aromatic fragrance, and soft texture, making it famous worldwide Research has shown that rice bran hydrolysates from Hom Mali white rice can improve cardiovascular health by reducing oxidative stress, inflammation, dyslipidemia, insulin resistance, and hypertension Sung Yod Patthalung, a traditional red violet pigment rice variety cultivated in southern Thailand for over a century, is rich in fiber and vitamins, promoting better bodily functions and overall health.
E, protein, iron, phosphorus than others, which can make people who usually have its benefit on the blood circular, on the body of keeping fit , help to protect the forgetfulness diseases, and with anti-oxidance as well as oryzanol and gamma aminobutyric acid[3] It is very popular among consumer who appreciate its splendid flavor and certain nutrients for health benefits When these two best quality Thai rice varieties hybridization produce a new variety of rice is originated with the vivid colour of red ruby, flavorful soft texture and prominent nutritional property
1.1.1 Important of Tubtim Chum Phae rice
Tubtim Chum Phae rice is a nutritious variety of brown rice characterized by its red membrane, which is rich in antioxidants and has a low amylose content of 12.63% This rice is packed with health-promoting nutrients, including high levels of polyphenols, phenolics, and flavonoids It serves as an excellent source of manganese and provides significant amounts of selenium, phosphorus, copper, magnesium, and niacin (vitamin B3), along with various B vitamins such as B1, B2, B6, and essential acids like pantothenic acid (vitamin B5), para-aminobenzoic acid (PABA), folic acid (vitamin M), and phytic acid.
Tubtim Chum Phae rice is a gluten-free, high-fiber food that can be easily incorporated into various dishes Regular consumption of this rice may lower the risk of myocardial infarction and improve cholesterol levels in individuals with dyslipidemia by enhancing low-density lipoprotein (LDL) and high-density lipoprotein (HDL) concentrations Additionally, it offers antioxidant and anti-inflammatory benefits, reduces cancer risk, supports brain health against neurodegenerative disorders, and helps protect the body from ultraviolet radiation and pathogens.
1.1.2 Life cycle of Tubtim Chum Phae rice
Figure 1.1 Growth stages of the rice plant Part of the image collection of the
International Rice Research Institute (IRRI)
The Tubtim Chum Phae rice variety has a life cycle of approximately 130 to 144 days, influenced by environmental factors Its growth can be divided into three key phases: the vegetative phase, which spans from seed germination to panicle initiation; the reproductive phase, from panicle initiation to anthesis; and the ripening phase, which extends from anthesis to full maturity.
The vegetative phase of plant growth is marked by the emergence of tillers and an increase in leaf count, leading to a gradual rise in height This stage typically lasts between 55 and 85 days, beginning with seed germination and continuing until tillering is complete The initial part of this phase is known as the seedling stage, which starts immediately after germination.
The initial growth stage of a plant begins with the emergence of the first root and shoot, lasting until just before the first tiller appears During this phase, seminal roots develop alongside up to five leaves, with two additional leaves forming as the seedling grows Leaves continue to develop at a rate of one every 3–4 days in the early stage The late vegetative phase commences with tillering, continuing until the maximum number of tillers is achieved As the tillering stage progresses, the stem lengthens and growth in height ceases just before panicle initiation, marking the conclusion of the vegetative phase.
The reproductive phase of rice begins with the 'booting' stage, where the leaf stem swells to conceal the developing panicle Once the tip of the panicle emerges, the rice enters the 'heading' stage, during which the panicle becomes fully visible Following this, flowering occurs, starting a day after heading, and lasts for about seven days, allowing for pollination through the shedding of pollen The ripening phase commences with flowering and typically lasts around 30 days, concluding when the grains are mature and ready for harvest This phase includes several stages: milky, dough, yellow, ripe, and maturity, which are determined by the texture and color of the grains The duration of ripening varies by rice variety, ranging from 15 to 40 days, and is influenced by temperature, taking about 30 days in tropical climates and up to 65 days in cooler temperate regions.
Microbiota
Microbes are essential for sustaining life on Earth, yet our understanding of most microbial communities in various environments, including soils, oceans, and our own bodies, remains limited These communities play a vital role in biogeochemical processes, agriculture, biotechnology, and human health The term "microbiome" can be defined in two ways: as the collective genomes of microorganisms in a specific habitat or as the total set of microorganisms present in that environment.
The microbiome refers to the diverse communities of microorganisms, including bacteria, fungi, protozoa, archaea, and viruses, that inhabit various environments and even differ among individuals These microbial communities are dynamic, influenced by factors such as diet and climate, and maintain a symbiotic relationship with their hosts, which is essential for both parties In humans, the microbial population vastly outnumbers human cells, highlighting the importance of understanding these microorganisms for insights into human biology, drug responses, and disease susceptibility Similarly, in plants, the extensive diversity of microbes associated with roots, often referred to as the plant's second genome, plays a critical role in plant health and fitness, with interactions shaped by plant, microbial, and environmental factors.
Microbial communities are essential for plant health, significantly impacting their physiology and development Numerous studies have shown that microorganisms associated with plants can greatly influence seed germination, seedling vigor, growth, nutrition, disease resistance, and overall productivity Specifically, the plant microbiota consists of diverse microbial communities that inhabit and interact with various plant tissues, including roots, shoots, leaves, flowers, and seeds The diversity of microbes found in plant roots is particularly vast, numbering in the tens of thousands.
The complex microbial community associated with plants, often referred to as the plant's second genome, plays a vital role in plant health and global biogeochemical cycles Plants release 5 to 20% of their photosynthetic products into the rhizosphere, a microbially diverse region enriched by root exudates, contributing significantly to soil health Additionally, plants emit substantial amounts of methanol and isoprene into the atmosphere each year, which serve as potential carbon and energy sources for microorganisms By manipulating the plant microbiome, it is possible to decrease plant disease incidence, enhance agricultural productivity, minimize chemical inputs, and lower greenhouse gas emissions.
The interactions between plants and their microbiomes are intricate and dynamic, with the plant immune system playing a crucial role in shaping microbiome structure Plants generate a diverse array of antimicrobial compounds, such as phenolics, terpenoids, and alkaloids, both continuously and in response to pathogens The microbiota can be seen as an extension of the host genome, and microbiomes occupying the same niche in different plants can exhibit significant differences, especially when examined at detailed taxonomic levels like genus, species, and strain.
Plants have evolved alongside microbial symbionts, including bacteria, archaea, fungi, and protista, forming a close association that enhances plant fitness through essential genetic contributions This relationship positions plants as superorganisms, integrating selected symbiont microbiota with host cells to function as a cohesive unit The interactions between these partners are characterized by a complementarity of metabolic capabilities, particularly evident in rice plants.
Research indicates the presence of three distinct root compartments: the endosphere (root interior), rhizoplane (root surface), and rhizosphere (soil near the root surface), each hosting unique microbiomes with diverse communities of eubacteria and methanogenic archaea These microbial communities are influenced by factors such as geographical location, soil type, host genotype, and cultivation practices The colonization patterns of root-associated microbiomes across these niches suggest a rapid acquisition from soil and support a multistep model of microbiome assembly, where each niche plays a selective role Environmental factors, including temperature, pH, and chemical signals from bacteria, plants, and nematodes, dynamically shape the rhizosphere microbiome, facilitating selective associations between plants and their microbiomes As land plants thrive in soil rich in microbial diversity, they and their associated microbes have evolved to leverage this relationship for mutual benefit, with microbes converting critical nutrients into usable forms for plants Additionally, bacteria in the rhizosphere utilize carbon metabolites from plant root exudates, while beneficial soil microbes enhance pathogen resistance, improve water retention, and promote growth through hormone synthesis.
Microbiome engineering aims to manipulate the plant microbiome to enhance specific functions, such as reducing disease susceptibility, improving nutrient availability, increasing abiotic stress tolerance, and boosting crop yields A promising strategy involves leveraging the natural communication channels that have evolved between plants and their microbiomes Every part of a plant is inhabited by diverse microorganisms, including bacteria, archaea, and fungi, collectively referred to as the phytomicrobiome By understanding these interactions, we can optimize plant health and productivity.
The phytomicrobiome consists of microorganisms categorized as endophytic (inside plant parts), epiphytic (on aboveground plant parts), or rhizospheric (in soil near roots) These microorganisms are essential to plant health, forming a complex relationship with their host that is often described as a metaorganism or holobiont The composition and biomass of the microbiome significantly influence the interactions between plants and their surrounding environments.
The relationship between plants and microbes is crucial for plant health, productivity, and overall condition, with interactions ranging from beneficial to pathogenic The outcome of these interactions can vary based on plant species and nutrient conditions The aim of plant–microbiome engineering is to enhance beneficial outcomes, leveraging microbial functions such as nutrient cycling, soil organic matter mineralization, disease resistance, and resilience to abiotic stresses like drought and salinity These interactions are complex and influenced by factors such as plant species, soil type, and environmental conditions, including biotic and abiotic stresses, climate, and human activities Various soils and environmental stresses can trigger distinct physiological responses in plants, leading to different exudation patterns.
1.2.4 Microbiota application for plant growth promote
In recent years, plant-associated microbial communities have gained significant attention for their role in enhancing crop productivity and stress resistance, similar to the benefits achieved through traditional plant breeding It is widely acknowledged that all plants, along with nearly all their tissues, host diverse microorganisms that provide various advantages, such as improving nutrient uptake, preventing pathogen attacks, and promoting plant growth in challenging environmental conditions.
Microorganisms provide essential shelter and a carbon-rich food supply, with mycorrhizal fungi and root-nodulating bacteria being the most prominent examples that enhance nutrient uptake and fix nitrogen Numerous novel plant growth-promoting microorganisms (PGPM) are continually being discovered, which help plants resist stress through various mechanisms Symbiont-based strategies are being explored to improve crop resilience to abiotic stresses, particularly drought, a significant challenge for global agriculture Future research should leverage advanced genomic technologies to identify and select beneficial symbionts and their functional roles in promoting plant growth Additionally, a reevaluation of current agronomic practices is necessary, considering the influence of microbial communities on plant traits The article advocates for closer collaboration between traditional breeding programs and microbial research to enhance crop productivity, emphasizing the complex interplay between plant genetics and microbial community dynamics.
Plant-growth promoting microbes enhance the growth and resilience of various plant species, providing improved tolerance to multiple environmental stresses simultaneously.
Types of soil sample
Chemical soil quality is influenced by its mineral composition, organic matter, and environmental factors, and can become contaminated through natural processes or human activities In agricultural practices, chemical fertilizers are commonly used For instance, Chemical 1 incorporates green manure along with N-P-K fertilizer (16-8-8) at a rate of 25 kg per 1.6 m², while Chemical 2 combines green manure with N-P-K fertilizer (15-15-15) at 35 kg per 1.6 m² Additionally, Chemical 3 utilizes green manure and N-P-K fertilizer (16-8-8) at a similar application rate of 25 kg per 1.6 m².
Intensive Chemical soil refers to soil samples that have undergone significant chemical treatment, specifically with Intensive Chemical 1, which utilizes N-P-K fertilizer (15-15-15) at a rate of 50 kg per 1.6 m² This method also includes the application of insecticides twice and liquid bio-fertilizer three times, particularly after the rice harvesting process.
In corn planting, Intensive Chemical 1 employs a variety of insecticides and chemical fertilizers, utilizing urea at a rate of 12.5 kg per 1.6 m² and N-P-K (15-15-15) fertilizer at 50 kg per 1.6 m², both applied once Intensive Chemical 2 follows a similar approach, using herbicides and insecticides alongside urea at 20 kg per 1.6 m² and N-P-K (15-15-15) fertilizer at 20 kg per 1.6 m², also applied once.
Transition type soils are those that have shifted from chemical fertilizers to exclusively organic fertilizers In Transition 1, insect dung is applied at a rate of 30 kg per 1.6 m² Transition 2 involves the use of photosynthetic bacteria administered once Transition 3 incorporates chicken dung at 150 kg per 1.6 m², along with monthly applications of fermented banana shoots and photosynthetic bacteria.
Objectives
The purposes to study are:
1.4.1 To study and point out the effect of soil sample groups to growth and health, phenotype and microbiome system of rice plant
1.4.2 To study and evaluate soil types and environment for the development of the best rice plants
METHODOLOGY
Equipment and materials
Table 2.1 Equipment used for studies
No Name of device Type Country
10 Incubator chamber VWR / Shel Lab 5216 US
13 Deep freezer VXE 380 Czech Republic
1 Tubtim Chum Phae rice from 25 Moo 11 Ban Pongchueag, Namakhuea, Sahadsakhan, Kalasin 46140, Thailand
2 Soil samples, soil standard, cow dung, rice bran, seed, pot
3 Small glass bead used for cell lysis and big glass bead used for release cell from soil
4 Laboratory instruments such as a micropipette,15ml and 50ml falcon tube, Eppendorf tube, test tube rack, beaker, measuring cylinder, stirring bar, pipette tip, weighing paper, forceps, facemask, glove
5 The chemical used in the experiment: 0.85% NaCl, 1M Tris-HCl, 1M EDTA, 1.5M NaCl, DI water, 10% SDS, capsule of activate charcoal were used for release DNA, 3M sodium acetate (pH:5.2), absolute ethanol, 30% PEG ( MW00) were used DNA clean soil chemical, phenol mixed with chloroform, isoamyl alcohol rate 25:24:1 were used for protein clean, chloroform, isoamyl rate 24:1 were used for phenol clean, isopropanol was used for DNA precipitate, ethanol 70% was used for wash DNA, magnetic, nuclease- free water for dissolve DNA
Methods
Table 2.3 Data of soil sample collected from somewhere in Thailand
Name Address Name of owner Time for grow rice
Start organic rice farm for
Start organic rice farm for
Start organic rice farm for
Malai Tongjarern Grow rice more than 20 years
Chuen Laokonka Grow rice more than 20 years C3 Bankok, Kokpochai,
Mali Nadee Grow rice more than 20 years I1 Bankok, Kokpochai,
Start rice farm for 8 years
Start rice farm for 8 years
Jiirapon Jaikachem Start rice farm for 8 years
Table 2.4 Weight amount of the soil sample in growth rice plant
Real weight soil for using
Real weight soil for using
Real weight soil for using
Weight soil samples and 500g of soil standard and 48g rice bran was mixed with cow dung in the bag and mix well together Then, pour into pot and added more
4 kg of soil standard and mixed continue when finished add water more into pot did it flood and put pots outside of balcony overnight
To prepare seeds for sowing, soak them in water for about 5 minutes After soaking, remove any hollow, diseased, or underdeveloped seeds that float to the surface, and retain only the viable seeds that settle at the bottom Drain the water and plant the collected seeds in pots, ensuring they are pushed gently into the soil During the germination process, cover the pots with a lid in the evening and remove it in the morning Add a small amount of water daily to keep the seeds moist without flooding them, which can cause rot It's important to use water that has been allowed to sit overnight to let any sediment settle.
To grow rice successfully, add water daily and after 3 to 4 days, observe the germination process, where you'll see the first small root and tiller along with two tiny leaves emerging from the rice seed After one week, carefully remove the germinated seeds, retaining only the strongest ones for further growth.
Plant 15 different rice seeds in individual pots and monitor their growth weekly Observe variations in plant shape, tiller development, and leaf color, documenting your findings with photographs each week.
Weekly observations of rice plants involved counting the number of tillers and photographing the plants alongside a ruler to measure their height Additionally, the color of the rice leaves was assessed each week The best and worst performing rice plants were identified within each group, and a comparative analysis was conducted among the selected plants across different groups.
2.2.5 Measure pH of soil sample
Measuring soil pH is crucial for plant growth and disease management, as it influences nutrient absorption by plant roots In acidic soils, essential nutrients like calcium, phosphorus, potassium, and magnesium may become unavailable, while basic soils hinder the absorption of copper, zinc, boron, manganese, and iron An improper pH can lock nutrients in the soil, leading to deficiencies, and exacerbate issues with diseases, insects, and weeds Soil pH indicates the level of acidity or alkalinity, defined chemically as the log10 of hydrogen ions (H+) in the soil solution, with a scale ranging from 0 to 14, where 7 is neutral.
A solution with a pH value above 7 is classified as basic or alkaline, while a pH below 7 indicates acidity Most plants thrive within an optimal pH range of 5.5 to 7.5, making it crucial for gardeners to monitor soil pH for healthy plant growth.
1 Sample collected include 27 soil sample with Chemical soil, Intensive Chemical soil Transition soil per pot into sterilized 50 ml tube
2 Added DI water with 10 mL per tube
3 Bring to vortexed shake mixture vigorously on 30 min
4 Centrifuge for 5 min at 8000 rpm
5 Tested - the first turned on your pH meter and remove the cap to expose the sensor completely in the solution
6 Record the reading on the meter
Inspection of quantity microbiome in Chemical, Intensive Chemical,
Weigh approximately 2 g of sample using a weighing paper Transfer the sample into
1 Added 2 g small glass beads and 2 ml DNA extraction buffer
2 Vortexed for 2 minutes with per tube and hold it upright
3 Added 1 ml DNA extraction buffer no add SDS and vortex for 2 minutes with per tube, hold it upright
4 Added 1 ml DNA extraction buffer no add SDS, no add activate charcoal and vortex for 2 minutes with per tube, hold it upright
5 Centrifuge for 15 minutes at 8000 rpm and collect supernatant transfer into 15 mL tube Centrifuge again 5 minutes at 8000 rpm and collected supernatant
6 Added 800 àl 3M sodium acetate and 4 ml absolute ethanol
8 Centrifuge for 20 minutes at 8000rpm and collected pellet
9 Re-suspend the pellet with 500 àl DI water
10.Added 500 àl Phenol: Chloroform: Isoamyl alcohol (25:24:1), centrifuge for
2 min at 13500 rpm and collected equeous phase Collected the first time about
400 àl and the second time about 350 àl In this step, done for 2 time
11.Added 1 ml Chloroform: Isoamyl alcohol (24:1), centrifuge for 2 minutes at
13500 rpm and collected equeous phase In this step, done for 3 time
12.Added 800 àl 3M sodium acetate and 4 ml absolute ethanol
14.Centrifuge for 10 minutes at 13500 rpm and collected pellet
15.Washed with 1ml 70% ethanol, centrifuge for 2 minutes at 13500 rpm and collected pellet In this step, done for 2 time
16.Waited ethanol in sample dry at room temperature
17.Dissolved in 20 àl Nuclease – free water
Measure Nanodrop
The NanoDrop microvolume sample retention system by Thermo Scientific utilizes fiber optic technology and natural surface tension to effectively capture tiny sample volumes without the need for traditional containers like cuvettes or capillaries This innovative system features shorter path lengths, enabling a wide range of nucleic acid concentration measurements and eliminating the necessity for dilutions By minimizing the sample volume required for spectroscopic analysis, it allows for the integration of additional quality control steps in various molecular workflows, enhancing efficiency and boosting confidence in downstream results.
1 The first, clean the upper and lower optical surfaces of the microvolume spectrophotometer sample retention system by pipetting 1 àL of clean deionized water onto the lower optical surface
2 Closed the lever arm, ensuring that the upper pedestal comes in contact with the deionized water Lift the lever arm and wipe off both optical surfaces with a clean, dry, lint-free lab wipe
3 Open the NanoDrop software and select the Nucleic Acid application Use a small-volume, calibrated pipettor to perform a blank measurement by dispensing 1 àL of buffer onto the lower optical surface Lower the lever arm and select "Blank" in the Nucleic Acid application
4 Once the blank measurement is complete, clean both optical surfaces with a clean, dry, lint-free lab wipe
5 Choose the appropriate constant for the sample that is to be measured
6 Dispense 1 àL of nucleic acid sample onto the lower optical pedestal and close the lever arm Because the measurement is volume independent, the sample
18 only needs to bridge the gap between the two optical surfaces for a measurement to be made
7 Select "Measure" in the application software The software will automatically calculate the nucleic acid concentration and purity ratios Following sample measurement, review the spectral output
8 The software will automatically calculate the nucleic acid concentration and purity ratios Following sample measurement, review the spectral image to assess sample quality
9 A typical nucleic acid sample will have a very characteristic profile
RESULTS AND DISCUSSIONS
The effect of the soil on the revolution of a rice plant with each Chemical,
In week 3, the tiller count of rice plants was initiated, and by week 5, comparisons were made across different soil groups However, the differences in rice plant growth at week 5 were minimal due to the rainy season, which caused flooding in the fields This excessive water is not conducive to optimal plant growth, as it hinders the transfer of essential nutrients from the soil to the plant's tillers and leaves Ideally, the soil should maintain a thin layer of moisture, approximately 1 inch deep, to support healthy rice development.
During the growth period of rice plants, particularly when they are producing tillers and leaves, a 2 cm layer of soil is crucial When the soil surrounding the roots is saturated with water, the plants may exhibit reduced growth, which can hinder their nutrient absorption Throughout the first five weeks, there was no significant change in the height or tiller count among the rice plants However, by week six, noticeable growth occurred, with significant changes in both height and tiller development At this stage, the best and worst rice plants were selected for comparison from the Chemical soil, Intensive Chemical soil, and Transition soil groups.
3.1.1 Comparison the different of rice plant about the color of rice leaves and the tiller number of rice plant in Chemical soil group
Table 3.1 Variation in the phenotype of rice plant in Chemical soil group after every week from week 5 until week 8
Time The best rice plant The worst rice plant
Figure 3.1 Comparison the average of the tiller number of rice plant in Chemical soil group from week 3 until week 8
The analysis of tiller numbers in rice plants across different chemical soil groups reveals distinct growth patterns In Chemical 1, tiller development remains consistent and stable week after week Conversely, Chemical 2 shows initial strong growth, outperforming the other groups until week 8, where tiller numbers plateau, mirroring week 7 In contrast, Chemical 3 initially exhibits lower tiller counts but experiences a significant surge in week 8, ultimately achieving the highest tiller numbers, although fluctuations in growth are notable when observing the error bars.
Week 3 Week 4 Week 5 Week 6 Week 7 Week 8
Comparison the tiller number of rice plant in Chemical group
3.1.2 Comparison the different of rice plant about the color of rice leaves and the tiller number of rice plant in Intensive Chemical soil group
Table 3.2 Variation in the phenotype of rice plant in Intensive Chemical soil after every week from week 5 until week 8
Time The best rice plant The worst rice plant
Figure 3.2 Comparison the average of the tiller number of rice plant in Intensive
Chemical soil group from week 3 until week 8
Figure 3.2 illustrates the tiller count of rice plants grown in Intensive Chemical soil While the calculation error bars indicate no significant changes in tiller numbers, Intensive Chemical treatments exhibited substantial variations Initially, rice plants under Intensive Chemical 1 showed poor growth, but by weeks 5 to 7, their performance improved, with the best results observed in week 7 Conversely, Intensive Chemical 3 demonstrated the highest tiller count by week 8 In contrast, rice plants in Intensive Chemical 2 exhibited consistent growth, particularly peaking at week 6, though they did not achieve the best results by week 8 Overall, Intensive Chemical 2 showed a steady increase in tiller numbers each week without any signs of stagnation.
After week 6, this plant shows significant decline, exhibiting signs of pathogens and numerous yellow leaves As the yellow leaves die off, new growth appears, resulting in many tillers; however, these tillers are noticeably smaller, thinner, and shorter compared to those of other plants.
Week 3 Week 4 Week 5 Week 6 Week 7 Week 8
Comparison the tiller number of rice plant in Intensive
3.1.3 Comparison the different of rice plant about the color of rice leaves and tiller number of rice plant in Transition soil group
Table 3.3 Variation in the phenotype of rice plant in Transition soil after every week from week 5 until week 8
Time The best rice plant The worst rice plant
Figure 3.3 Compared average of the tiller number of rice plant in Transition soil group from week 3 until week 8
Figure 3.3 illustrates the tiller count of rice plants in transition soil The analysis revealed some calculation errors indicated by error bars during the initial weeks, although there was minimal change observed by the end of the week.
Between weeks 6 and 8, significant changes were observed in the growth of rice plants across the three transitions Transition 1 showed a stable tiller count initially, but from weeks 7 to 8, growth stagnated due to limited space and insufficient fertilizer in the pot In contrast, Transition 2 performed poorly, as it relied solely on photosynthetic bacteria without adding any nutrients, resulting in minimal soil fertility and inadequate development Transition 3, however, continued to thrive, albeit at a slower pace during weeks 7 and 8, thanks to the regular application of organic fertilizers such as chicken dung and fermented banana shoots, which enriched the soil with essential nutrients.
Week 3 Week 4 Week 5 Week 6 Week 7 Week 8
Comparison the tiller number of rice plant in
3.1.4 Comparison the different of rice plant between Chemical soil, Intensive Chemical soil, Transition soil groups
Table 3.4 Variation in the phenotype of rice plant between three soil groups after every week from week 5 until week 8
Time The best rice plant The worst rice plant
Figure 3.4 Comparison the average of the tiller number of rice plant in Chemical soil, Intensive Chemical soil, Transition soil group from week 3 until week 8
Figure 3.4 Shows the tiller number of rice plant between Chemical group,
During the growth of rice from week 3 to week 5, the tiller development in all soil groups remained relatively unchanged due to excessive flooding during the rainy season, which caused the rice plants to exhibit minimal growth Notably, significant changes in tiller numbers were observed only in week 6 The Intensive Chemical and Chemical soil groups demonstrated superior tiller development; however, the Intensive Chemical group showed signs of pathogen infection, resulting in smaller, thinner plants with yellowing leaves In contrast, the Transition group, which lacked synthetic fertilizers and pesticides, performed the poorest due to the absence of essential nutrients, highlighting the importance of chemical fertilizers like N-P-K and liquid bio-fertilizers in promoting healthy rice growth.
21 days 28 days 35 days 42 days 49 days 56 days
Comparison the tiller number of rice plant in three soil groups
To maintain soil health and provide essential nutrients for crops, it is important to implement practices such as crop rotations, utilizing crop residues, animal manures, legumes, green manures, off-farm wastes, and biological pest control Additionally, regular soil testing every three months is crucial to ensure the appropriate application of fertilizers, as inadequate soil sampling can lead to insufficient nutrient levels for rice cultivation.
3.1.5 Observation of the change of pH in three types of soil sample
Table 3.5 Calculation the average of pH all of soil sample to observe the different of soil to rice plant
Average pH of soil sample
Soil sample pH of soil
Before rice plant Week 5 Week 8
Figure 3.5 The average of pH of soil sample during growth rice progress and compare the differences between each sample
Measure pH soil from rice field soil from pot after 5 weeks soil from pot after 8 weeks
The study of soil pH in rice fields reveals significant insights into soil properties and plant health After analyzing soil samples from various treatment groups, it was found that the average pH of the rice field remained relatively stable, indicating no significant changes However, the Intensive Chemical treatment group exhibited considerable fluctuations by week 5, coinciding with visible signs of disease in the rice plants, such as yellowing leaves Measurements taken at week 5 showed that the optimal pH for rice growth was around 6, which enhances nutrient availability, particularly during the rainy season when flooding occurs By week 8, a decline in soil pH to levels between 4-5 indicated highly acidic conditions, potentially leading to toxic concentrations of soluble aluminum, iron, and manganese detrimental to plant growth Given that the pH scale is logarithmic, even slight changes in pH can significantly impact rice plant health.
The result of Nanodrop
In a study involving twenty-seven soil samples, researchers cultivated rice plants and monitored their growth weekly They compared the development of the rice plants to identify the most robust specimens within each large group Subsequently, the best rice plants from these groups were selected, and soil samples were collected for DNA extraction to analyze the DNA concentration present in the soil.
Table 3.6 The result of Nanodrop after DNA extraction
Table 3.2 presents the Nanodrop analysis results for DNA concentration in soil samples Most samples exhibited relatively high DNA concentrations; however, the C2/1 and T3/1 samples showed notably low levels, while C2/3 and I2/3 had very minimal DNA presence The Nanodrop measurements indicated that the concentration of DNA in these samples was nearly absent Additionally, all samples demonstrated contamination with other chemicals, as reflected in the A260/280 and A260/230 purity ratios.
The result of DNA extraction for check microbiome from soil samples
After checked concentration of DNA by Nanodrop technique continues with electrophoresis
Product for extract DNA was been checked by gel electrophoresis is gel 0.8 % agarose in 50 ml 1X TAE buffer The result of electrophoresis showed in this picture 3.9
Figure 3.6 The result of electrophoresis of DNA from soil samples of the best rice plant Land M: 1 Kb DNA ladder, land 2: C2/1, land 2: C2/2, land 3: C2/3, land 4: I2/1, land 5: I2/2, land 6: I2/3, land 7: O3/1, land 8: O3/2, land 9: O3/3
Figure 3.6 Shows the result of the concentration of DNA inside soil samples
There was appeared light DNA on each run line of land 4, land 5, land 8, land 9 so DNA extraction succeed and observed smear on land 2, land 6 but with land 1, land
Lands 3 and 7 showed no significant observations due to the chemical composition of soil in land 1 and land 3, which contains high levels of chemicals, making DNA extraction challenging Additionally, land 7, known for rice cultivation, consistently produces poor rice plants, indicating a lack of microbiome diversity Unfortunately, there was insufficient time to conduct further molecular biology analyses.
CONCLUSIONS AND SUGESTIONS
- No different of tiller number of rice but rice show different character about height and color of rice leaves in each group of soil
- Observation some disease in Intensive Chemical 3
- Rice plant of Chemical soil group is the best rice plant in three groups
- Water, fertilizer and space very important of rice plant
1 Rice was observed only 8 weeks and it should be observing to flower and ripening phase
2 Repeat experiment with better environment condition for growing rice plant
4 Should be develop the protocol for DNA extraction with Chemical soil group After success should be purify and sequence
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