The present Thai and world populations have been stepping into an aging society. To keep the elderly in good health and protected by diseases such as diabetes, cancer cardiovascular disease caused by fat blockage, hypertension etc. is the important problem. One way to solve this dilemma is a healthy diet which is eating safe and organic food without contaminations. Highly fragrant rice containing beneficial nutrients i.e., anthocyanin and oryzanol helps to reduce obesity and blood sugar level. The most and highly nutritious rice are colored black to red. In addition, Thai rice production uses harmful chemicals which is another problem that directly affects producers, consumers, and the environment. These significant problems were recognized by the research team. Thus, there is a need for breeding of rice with high nutrients, resistant to diseases and insects that can organically grow to make it suitable to consume by elderly and people in healthy diet. Also, reducing the use of chemicals to be safe for producers and consumers as well as being environmentally friendly needs to be integrated.
Project 1 Yield Trials, Plant Protection and Seed Production of Non-photoperiod Sensitive, Semi-dwarf, Aromatic, Red, Non-glutinous/Glutinous and High Nutritional Value of Sang Yod Phatthalung Rice Lines.
The improvement of Sang Yod Phatthalung rice varieties to become non-photoperiod sensitive, semi-dwarf, aromatic, red, non-glutinous/glutinous and high nutritional value was conducted using hd1, hd2, hd4, hd5, sd1, fgr, wx and SSIIa markers by molecular marker-assisted backcrossing (MAB). Presented non-glutinous/glutinous, semi-dwarf, early flowering and aromatic rice can be grown throughout the year. Sang Yod Phatthalung with non-glutinous and glutinous rice for two new varieties will be registered by PVP law, plant species royal decree act of 1999 in 2020 budget. Data for growth characteristics of 8 rice lines of non-glutinous and 2 rice lines of glutinous were collected, including the full tillering stage (i.e. leaf hair, shape of the tongue leaf and leaf cladding color), flowering phase (i.e. stem diameter, banded, 50% flowering days, color of the female stamens, plant height, number of panicles, corner of the flag leaf and emergence of the sheaves), maturity (i.e. seed loss, sheaves, harvest date and length of the panicles) and after harvest (i.e. paddy weight of 1000 grains, paddy color, type of grain, brown rice shape, amylose content, gelatinization temperature, gel consistency, aroma and resistance to disease and pests). Pure lines were produced for two seasons; rainy season 2019 and dry season 2020. Sang Yod Phatthalung with non-glutinous rice showed 75% of flowering days in 113 and 114 days, while the number of anticipated breeder seeds were selected with 53 and 130 plants, respectively. Sang Yod Phatthalung with glutinous rice showed 75% flowering days in 102 and 99 days while the number of anticipated breeder seeds were selected with 35 and 123 plants, respectively.
Moreover, Sang Yod Phatthalung with non-glutinous and glutinous rice was grown in rainy season 2019 at Maejo University organic farm in accordance with USDA and IFOAM standard. The yield of Sang Yod Phatthalung with non-glutinous/glutinous showed 309-647 kg/rai. Rice shapes including paddy width, paddy length, paddy thickness, width of the grain brown rice, length of brown rice seeds and thickness of the grain brown rice were 1.70-1.80, 8.30-9.80, 1.60-1.70, 1.70-1.80, 5.90-6.80 and 1.40-1.60 mm., respectively. As for the yield in the dry season 2019, the milling characteristics of Sang Yod Phatthalung rice seed varieties to become non-photoperiod sensitive, semi-dwarf, aromatic, red, non-glutinous/glutinous and high nutritional value were not chalky grain, with 68.61-85.48% of whole kernels and 14.95–31.77 % of broken rice. Physical characteristics of the seed, the length of the grain was between 5.79 - 6.69 mm. classified as short to class 2 long grain or long grain, and a slender grain shape with the ratio of length to width between 3.64 - 4.10 mm. In terms of chemical characteristics, the flow gel distance was between 27.83 - 84.17 mm and classified as hard to soft-cooked flour. Elongation rate of cooked rice was between 1.33 - 1.54. The percentage amylose content was 3.7 - 23.35 %, classified as glutinous rice and medium amylose non-glutinous rice. The decomposition value in a 1.7% KOH alkali solution was between 2 – 7, categorized as in low to high decomposition level. In addition, it contains aromatic substances (2AP) between 1.16 - 3.17 ppm.The nutritional quality revealed 0.29-0.39 mg of vitamin E, 0.34-0.41 mg of vitamin B1, 1.31-1.72 mg of iron, 0.39-0.96 mg of zinc, 0.14-0.24 mg of copper, 31-53 mcg of folate and 618.08-802.54 mg/kg of gamma oryzanol whereas no total anthocyanin (peonidin), carotenoid lutein and beta-carotene were detected. Moreover, Sang Yod Phatthalung with non-glutinous/glutinous rice displayed a total antioxidant active (ORAC) at 13,294.19-17,0330 ?moles TE, total antioxidant active (FRAP) at 2,507.09-3,516.61 ?moles TE, total polyphenol at 384.84-565.69 mg eq GA, total anthocyanin (cyanidin, peonidin) at 7.91-12.99 mg and total anthocyanin (cyanidin) at 7.91-12.91 mg.
The registration of the new plant variety, in terms of selection, reservation and collection for local and forest plants, will be filed according to Section 52 of Plant Variety Protection Act B.E. 2542. Moreover, the agreement with Plant Variety Protection Division, Department of Agriculture was made regarding the benefits shared from Sang Yod Phatthalung with non-photoperiod sensitive, semi-dwarf, aromatic, red, non-glutinous rice (namely Maejo 23) and Sang Yod Phatthalung with non-photoperiod sensitive, semi-dwarf, aromatic, red, glutinous rice (namely Maejo 24). It was successfully completed until the end of the procedure. Successively, the fifth step to apply for the registration of a new plant variety and to apply for certification of registered plant variety with Plant Variety Protection Division, Department of Agriculture was made. In which, the characteristics of the variety will be used for the submission of Sang Yod Phatthalung with non-photoperiod sensitive, semi-dwarf, aromatic, red, non-glutinous rice (namely Maejo 23) were as follows; semi-dwarf, erect clump, diameter of the stem was 4.16 mm, height of the trunk to the rice neck was 66 cm., leaf blade: pubescence of surface some weak, cleft leaf: shape of ligule cleft , green leaf sheath, white stamens and corner of the flag leaf was erect, 50% flowering days in dry season was 118 days and 111 days in rainy season. A lot of panicle base showed 16 panicles per plant. Harvest maturity in dry season was 148 days and 141 days in rainy season. Length of panicles was 27.65 cm. Paddy was straw color. Mean of shape including width of paddy, length of paddy, thickness of paddy, width of the grain brown rice, length of brown rice seeds and thickness of the grain brown rice were 9.90, 2.00, 1.70, 6.55, 1.65 and 1.53 mm., respectively. Seeds was slender (length-to-width ratio was 3.97). One thousand grain weight (14% moisture) was 17 g. Number of spikelets per panicle was 182 seeds, seed set was 82 % and seed loss was 6-25 %. Amylose content was 11.55% and with low gelatinization temperature. Seed decomposition value (1.7%KOH) was 7.0. Concentration of 2AP was 2.77 ppm. Sang Yod Phatthalung with non-photoperiod sensitive, semi-dwarf, aromatic, red, glutinous rice (namely Maejo 24) characteristics that will be used for registration were as follows; semi-dwarf, erect clump, diameter of the stem was 4.71 mm., height of the trunk to the rice neck was 62 cm., leaf blade: pubescence of surface some weak, cleft leaf: shape of ligule cleft , green leaf sheath, white stamens and erected corner of the flag leaf. Fifty percent flowering days in dry season was 99 days and 104 days in rainy season. A lot of panicle base showed 16 panicles per plant. Harvest maturity in dry season was 129 days and 134 days in rainy season. Length of panicles was 24 cm. Paddy was straw color. Means of shapes including width of paddy, length of paddy, thickness of paddy, width of the grain brown rice, length of brown rice seeds and thickness of the grain brown rice were 10.00, 2.05, 1.70, 6.65, 1.70 and 1.50 mm., respectively. Seed was slender (length-to-width ratio was 3.91). A thousand grain weight (14% moisture) was 17 g. Number of spikelets per panicle were 219 seeds with 84 % seed set and 6-25 % seed loss. Amylose content was at 6.48% and with low gelatinization temperature. Seed decomposition value (1.7%KOH) was 7.0. Concentration of 2AP was 2.84 ppm. Breeder seeds were produced for this season. Sang Yod Phatthalung with non-photoperiod sensitive, semi-dwarf, aromatic, red, non-glutinous rice (namely Maejo 23) showed 75% of flowering days in 108 days, while a number of breeder seeds were selected with 142 plants and Sang Yod Phatthalung with non-photoperiod sensitive, semi-dwarf, aromatic, red, glutinous rice (namely Maejo 24) showed 75% of flowering days in 95 days, while a number of breeder seeds were selected with 214 plants.
Project 2 Sub-project 2 Selection, Observation and Yield Trials of Non-photoperiod Sensitive, Semi dwarf, Aromatic Non-Glutinous, Resistant to Diseases and Insect and High Nutritional Value of Hom Mali Dang Rice Lines
Nowadays, a trend of rice with high nutritional value or with special properties has continuously increasing. Therefore, such rice varieties should be studied and selected,to cultivate and increase productivity for exportation by employing marketing strategies in terms, special properties, and distinct packaging of public interest. Thus, this project aims to select, study, and yield test of non-photoperiod sensitive, semi dwarf, aromatic non-glutinous, resistant to diseases and insect, and high nutritional value of Hom Mali Dang rice lines. During the first year (2020), season 1 (rainy season 2019), an observation trial, planted 2 rows of 50 F7 lines non-photoperiod sensitive, semi dwarf, aromatic non-glutinous, resistant to diseases and insect and high nutritional value of Hom Mali Dang and the 16 plots with the highest yield potential were selected. It was found that the between yield was 341-775 kg per rai. Then, 298 plants with good agricultural characteristics were selected. Self-pollinated F8 seeds were produced and selected by molecular markers; Bph3, xa5, qBL1, qBL11. The most There were 105 lines with homozygous genotypes of the Bph3, xa5, qBL1, qBL11 gene were selected with the flowering date average of 75% in between 86-111 days. In season 2 (dry season 2020), the advance trial planted 4 rows of 35 F8 lines non-photoperiod sensitive, semi dwarf, aromatic non-glutinous, resistant to diseases and insect, and high nutritional value of Hom Mali Dang. and the 16 plots with the highest yield potential were selected. It was found that the between yield was 748-1,082 kg per rai with the flowering date average of 75% in 89-131 days. And 186 plants One hundred eighty-six lines with good agricultural characteristics were selected. As a result, the 16 lines of non-photoperiod sensitive, semi dwarf, aromatic non-glutinous, resistant to diseases and high nutritional value of Hom Mali Dang with the highest yield potential were selected and self-pollinated F9 seeds were produced. The, F9 seeds were then selected by molecular markers; qBL1, qBL11, fgr, Wx/wx, Wxa/Wxb, SSIIa, Rc, OsB1. It was found that all genotypes of the 16 lines were homozygous at qBL1, qBL11, fgr, Waxy, SSIIa, Rc, OsB1 genes. Thus, qBL1 and qBL11 genes control rice to have resistance to blast disease. The fgr gene controls the fragrant rice while waxy genes control non-glutinous/glutinous rice on which all 16 lines were all non-glutinous. SSIIa gene controls rice to have a low starch gelatinization temperature. The Rc and OsB1 genes control the rice to red and black, respectively. These studies of the genotyping were able to select rice that have properties of fragrant rice, low starch gelatinization temperature, red and black colors of rice with high nutritional value. Therefore, they can be planted and further tested on Intra-station trial.
Project 3 Study of Chemical and Milling Quality of Rice Varieties from Sub-project 1 and Sub-project 2
Milling, physical and chemical qualities are important for rice breeding to obtain the market-potential, healthy and processing suitable rice. 14 non-glutinous rice samples (3 reference samples and 11 breed samples) and 10 glutinous rice samples (2 reference samples and 8 breed samples) were used in this research and analyzed for milling, physical and chemical qualities. It was found that the moisture content of paddy and milled non-glutinous rice were in the ranges of 12.45-14.00 and 8.25-9.14, respectively. The moisture content of paddy and glutinous rice were in the ranges of 12.05-13.20 and 7.99-9.04, respectively. The milled rice yield of non-glutinous rice and glutinous rice was 53.58-65.64 and 55.58-65.64 of the paddy rice, respectively. The total content of the whole and head rice of the non-glutinous and glutinous rice was perfectly good and had the values of 52.39-84.61 and 72.01-92.11 of the total milled rice, respectively. All rice samples had the long grain with the ratio of length/width higher than 3.0. 80% (8 samples) of the cooked starch obtained from the breed non-glutinous rice were hard. The 100% (3 samples) of the cooked reference non-glutinous rice were soft. For all of the cooked glutinous rice starch were soft. Breeding rice increased the hardness of the cooked non-glutinous rice starch. The elongation ratio of the non-glutinous rice was in the range of 1.35-1.54 while thet of the glutinous rice was in the range of 1.33-1.49. The amylose content of the non-glutinous rice was in the range of 9.96-23.35%. The breed non-glutinous rice samples were classified as low-medium amylose rice while the reference non-glutinous ones were classified as low amylose rice. For all glutinous rice were low amylose rice with 3.70-6.18% amylose contents. The degradation scores in the alkali of rice non-glutinous was 2-7 and that of the glutinous rice was 4-7. The pasting properties were as follows:- the gelatinization temperature of non-glutinous and glutinous rice flour was in the range of 86.75-91.51 and 73.99-79.33 ?C, respectively. The peak viscosity of non-glutinous and glutinous rice flour was 1340.00-4166.67 and 1534.21-2678.00 cP, respectively.
Project 4 Research project 4: Improvement of rice varieties to increase anthocyanins by molecular marker-assisted selection for increased nutritional values
Black rice has high accumulation of anthocyanins on pericarp tissues. OsB1 and OsC1 genes are important for the controls of anthocyanin biosynthesis in rice. OsDFR gene is structural gene that encodes the vital enzyme in anthocyanin biosynthesis pathway. Our research granted in the year of 2019 showed that DNA markers specific to OsB1, OsDFR and OSC1 genes could be used to distinguish between white and black rice. In this study, we used these DNA markers to study the F2 population of 300 plants derived from the cross between white pericarp rice (Pathumthani 1) as a receptor and black pericarp rice (Kham Noi) as a donor. The inheritance of DNA markers specific to OsB1, OsDFR and OSC1 genes was analyzed by Chi-square test with 1, 2 and 3 genes, respectively. The results showed that DNA markers specific to OsB1 and OsDFR followed the Mendelian manner but DNA marker specific to OsC1 did not. Analysis of anthocyanin content by pH Differential method in seeds of F2 progenies showed that plant no 299 had highest anthocyanin content of 35.04 mg/100 gDW, consistent with 39.52 mg/100 gDW of the cyanidin-3-O-glucoside (C3G) content determined by HPLC method. Antioxidant activity was analyzed by DPPH assay in seeds of F2 progenies and the result showed that plant no. 199 had highest antioxidant activity of 9.7 ?moles/gDW which was similar to that of Kham Noi parent. Analysis of inheritance of F3 pericarp colors by Chi-square test showed the ratio of 3 colored: 1 colorless, corresponding to genotype of OsB1 marker of 3 A_: 1 aa. The relationship between genotype of DNA markers specific to OsB1, OsDFR and OsC1 and phenotype of F2 population was analyzed by ANOVA and regression method. The results suggested that only OsB1 marker was related to anthocyanin content, antioxidant activity pericarp color which had R2 of 31.9 %, 40.6 % and 69.5 %, respectively.
The inheritance of DNA markers specific to OsB1, OsDFR and OSC1 genes was analyzed by Chi-square test with each gene in 321 plants of BC1F1 population. The results showed that only DNA marker specific to OsB1 followed the Mendelian law which had the ratio of 1:1 (Aa:aa). The 15 BC1F1 plants which were heterozygous for DNA markers of OsB1, OsDFR and OSC1 genes were selected. They had different pericarp color including 8 black, 4 dark brown, 2 dark red and 1 red which were backcrossed with Pathumthani 1 for production of BC2F1 seeds and selfing for production of BC1F2 seeds. Antioxidant activity was analyzed by DPPH assay in seeds of BC1F1 and the result showed that plant no. 297 had highest antioxidant activity of 12.8 ?moles/gDW which was higher than 8.8 ?moles/gDW of Kham Noi parent.
In this study, the CAPS marker of OsB1 had relationship with phenotype of anthocyanin content, antioxidant activity and pericarp color. Therefore, it could be used for selection of rice with colored pericarp, high anthocyanin and antioxidant activity in the early stage of development which will be beneficial to plant breeding for facilitating selection and shortening the time in breeding Thai rice for high nutritional value. In addition, the seeds of BC2F1 and BC1F2 which were selected by the DNA markers and had black and red pericarp color, high anthocyanin and antioxidant activity, will be further used for improvement of rice varieties with increased nutritional value.
Project 5 Improvement of rice varieties to increase lutein or zeaxanthin by gene specific DNA marker selection
Lutein is the most abundant plant carotenoid that plays a role in retina protection against photo-stress. The aim of this study was to develop lutein or zeaxanthin specific gene markers from rice which, will be used in breeding program for developing high lutein or zeaxanthin varieties. Lutein content was examined by High Performance Liquid Chromatography. The result showed that Kum Noi, black pericarp rice, had the highest lutein content and was used as donor parent for F1, F2, BC1F1, BC1F2 และ BC2F1 populations production. Carotenoid biosynthesis gene sequence and expression level from different pericarp color rice were analyzed by transcriptome and whole genome sequencing using for gene specific marker development. There are 15 of gene specific marker were designed which were 11 genes of carotenoid synthesis pathway (PSY1 /crtB; phytoene synthase, PDS; phytoenede saturase, ZISO; ?-carotene isomerase, ZDS ; ?-carotene desaturase, CrtISO; carotenoid isomerase, lcyE; lycopene ?-cyclase, lcyB; lycopene ?-cyclase, LUT5/CYP97A3 ; cytochrome P450-type hydroxylase A3, Lut1, CYP97C; cytochrome P450 carotene epsilon-hydroxylase , CYP97B2; cytochrome P450 B and OsEHY /HYD3; ? -carotene hydroxylase), 1 SET DOMAIN gene ( SDG8; histone methyltransferase), 1 bHLH transcriptional factor gene (bHLH_B2_Kala4) and 2 of cleavage genes (LCD_CCD1; lycopene cleave dioxygenase and NCED2;9-cis-epoxycarotenoid dioxygenase 2/ CCD4b; carotenoid-cleavage dioxygenase 4b/Zeaxanthin cleavage oxygenase). Then simple regression analysis revealed that B2_Kala4 showed highest association with lutein (17.13%) and LCD_CCD1 showed highest association with zeaxanthin (1.73%). These gene specific markers will be used in molecular marker assisted selection for high lutein breeding.
Project 6 Improvement of rice variety to increase resistant starch using molecular marker-assisted selection
Resistant starch is beneficial for comsumer’s health. This research aimed to improve rice variety and develop molecular marker linked to resistant starch content in rice. The correlations between resistant starch content and molecular marker specific to SSIIIa, Waxy and SBEIIb gene which reported affecting on resistant starch content were tested in F2 population from two crossing, RD-Maejo 2 x RD 43 which are glutinous and non-glutinous rice respectively, and Pathum Thani 1 x Chao Lueang which are both non-glutinous rice. The results showed that the resistant starch content of 100 F2 samples from RD-Maejo 2 x RD 43 significantly correlated with marker specific to Waxy gene (r = 0.704, p < 0.000) and non-glutinous rice contained more resistant starch content than glutinous rice. Likewise, the resistant starch content of 92 F2 samples from Pathum Thani 1 x Chao Lueang significantly correlated with marker specific to Waxy gene (r = 0.682, p < 0.000). Rice with high amylose allele from Chao Lueang had more resistant starch content than rice with low amylose allele from Pathum Thani 1. The results indicated that the amylose content was positively related to the resistant starch content in rice. In addition, 100 seeds of BC1F1 were produced.
Project 7 Isolation of rice pathogens and rice endophytic bacteria from organic and chemical rice farms in northern part of Thailand for biological control of organic rice diseases
Rice bacterial leaf blight and rice blast caused by Xanthomonas oryzae pv. oryzae and Pyricularia oryzae are the major problem for rice production in Thailand. Chemical control was shown to be less effective. Endophytic bacteria are bacteria that have intimate relationship with their host without inducing any pathogenic symptom. Therefore, this study was focused on the biocontrol of rice pathogens and plant growth enhancement by endophytic bacteria from rice. These two rice pathogens were collected from various area in northern part of Thailand, particularly Chiangmai, Chiangrai, Lumpoon, Lumpang, Prae, Nan, Payao and Mae Hong Son. Eight strains of Xanthomonas oryzae pv. oryzae were obtained from Chiang mai and Payao area. Nineteen isolates of rice blast fungi, genus Pyricularia, were obtained from 8 provinces. From the rice pathogenicity test, Xoo and Pyricularia were 5 and 2 in severity, respectively.
Total of 471 endophytic bacteria were isolated from healthy rice stem and rice seed. The identification results show that Gram-negative bacteria were belonged to the group of Proteobacteria and Bacteroide and gram-positive bacteria were in the group of Firmicutes and Actinobacteria. Gram-negative endophytic bacteria were belonged to the genera of Acinetobacter, Agrobacterium, Burkhoderia, Citrobacter, Enterobacter, Klebsiella, Methylobacterium, Pantoea, Pseudomonas, Serratia, Sphingobacterium, Spingomonas and Stenotrophomonas, while gram-positive bacteria were in the genera of Bacillus, Brevibacillus, Curtobacterium Lysinibacillus, Paenibacillus, Streptomyces and Virgibacillus,. These bacteria were evaluated in vitro for inhibition of X. oryzae pv. oryzae by spot on lawn. Among these, it was found that fifty-five isolates comprising of Acinetobacter, Bacillus, Citrobacter, Klebsiella and Pseudomonas exhibited the antagonistic activity against X. oryzae pv. oryzae. Cell free supernatant of these antagonist bacteria were then investigated the antibacterial activity by agar well diffusion method. Only fourteen isolates of Pseudomonas and Bacillus expressed the inhibitory effect on X. oryzae pv. oryzae. The highest diameter of inhibition zone was obtained from Bacillus velezensis SK63-R283. These 471 bacteria were also tested against P. oryzae by dual culture assay. Total of 75 isolates comprising of the genera of Bacillus, Paenibacillus, Pseudomonas, Enterobacter and Klebsiella showed the inhibitory effect on fungal growth. Bacillus pumilus SK63-R260 showed the highest percentage of fungal growth inhibition at 65.57 %.
The plant growth enhancement of rice endophytic bacteria were investigated by their activities of phosphate solubilizing, nitrogen fixation and production of indole acetic acid. The phosphate solubilizing activity of bacteria were tested on National Botanical Research Institute’ phosphate growth medium (NBRIP) agar. It was found that 98 isolates showed the ability to solubilize phosphate according to clear zone around bacterial colony. They belonged to the genera of Acenitobacter, Bacillus, Brevibacillus, Enterobacter, Klebsiella, Paenibacillus and Pseudomonas. These bacteria were then carried out phosphate solubilizing activity in liquid medium. Results showed that Acinetobacter sp. SK63-R222 obtained the highest solubilizing ability of Ca3(PO4)2, about 261.66 ?g/ml. According to the growth in nitrogen free medium, sixty-six isolates belonging to Acinetobacter, Bacillus, Lysinibacillus, Paenibacillus and Staphylococcus, were able to fix nitrogen. All bacterial isolates were also screened indolic compound production by spectrophotometer. The result of quantitative analysis of IAA production from 77 bacterial isolates showed that Klebsiella pneumoniae SK63-R155 produced the highest IAA (237 ?g/ml).
In the study of rice disease control by biological methods, Bacillus velezensis SK63-R283 bacteria were selected for the pot experiment. It was found that Bacillus velezensis SK63-R283 could reduce the severity of bacterial leaf blight in rice and rice blast disease by 55% and 25%, respectively.
The optimized media were carried out to produce antibacterial substance against Xoo. Commercial bacterial culture media, comprising of nutrient broth, trypticase soy broth and potato dextrose broth, were investigated. The suitable medium for production cell mass and antibacterial substance by Bacillus velezensis SK63-R283 was trypticase soy broth.
The formulation of Bacillus velezensis SK63-R283 for controlling bacterial leaf blight were also studied. Bamboo biochar and kaolin were used as carriers in this experiment. These two substances were effective to maintain bacterial survival with 9 log CFU/g after 2-month storage. Pot experiments using seed treatment and foliar spray showed the decrease in bacterial leaf bight by 35%.
These results suggested that rice endophytic bacteria can be used for biological control of rice diseases and they can be potential for enhancing the plant growth. The others bioformulation of will be developed in ours research lab and the safety of this endophytic bacterium must be test before using in the rice field