Millet Varieties Research Articles

On-Farm Grain Yield Stability and Farmer Perceptions on Pre-release Pearl Millet Lines in Zimbabwe

Farmer participation in on-farm research does not only accelerate information gathering but also results in adoption of new research products. Here, we report on-farm trials conducted across five districts of Matabeleland province in Zimbabwe to explicate grain yield stability and farmer perceptions on eight pre-release pearl millet (Pennisetum glaucum L.) lines. The results indicated that genotypic effects on grain yield were significant in both individual and across-site analysis of variance and that farmers prioritize earliness and grain yield as must-have traits in millet varieties. The five districts were grouped into two distinct environments, with four districts (i.e., Bulilima, Gwanda, Matopos and Tsholotsho) in one group (i.e., DGrp1) and Mangwe district forming the second group (DGrp2). Pearl millet pre-release lines PM1 (1.425 kg ha-1), PM9 (1.043 kg ha-1) and PM6 (761.8 kg ha-1) showed high yield, stability and were the most preferred by farmers. In conclusion, on-farm trials may offer the quickest possible solution to boost low-pearl millet production resulting from the continuous use of unproductive landraces and old varieties by farmers.

Comparative Transcriptomic Analysis Reveals Domestication and Improvement Patterns of Broomcorn Millet (Panicum miliaceum L.).

Broomcorn millet (Panicum miliaceum L.) is one of the earliest crops, domesticated nearly 8000 years ago in northern China. It gradually spread across the entire Eurasian continent, as well as to America and Africa, with recent improvement in various reproductive and vegetative traits. To identify the genes that were selected during the domestication and improvement processes, we performed a comparative transcriptome analysis based on wild types, landraces, and improved cultivars of broomcorn millet at both seeding and filling stages. The variations in gene expression patterns between wild types and landraces and between landraces and improved cultivars were further evaluated to explore the molecular mechanisms underlying the domestication and improvement of broomcorn millet. A total of 2155 and 3033 candidate genes involved in domestication and a total of 84 and 180 candidate genes related to improvement were identified at seedling and filling stages of broomcorn millet, respectively. The annotation results suggested that the genes related to metabolites, stress resistance, and plant hormones were widely selected during both domestication and improvement processes, while some genes were exclusively selected in either domestication or improvement stages, with higher selection pressure detected in the domestication process. Furthermore, some domestication- and improvement-related genes involved in stress resistance either lost their functions or reduced their expression levels due to the trade-offs between stress resistance and productivity. This study provided novel genetic materials for further molecular breeding of broomcorn millet varieties with improved agronomic traits.

Response of pearl millet (Pennisetum glaucum) to crop residue application and potassium management options under rainfed condition

A field experiment was conducted during rainy (kharif) seasons of 2021 and 2022 at ICAR-Indian Agricultural Research Institute, New Delhi to study the impact of crop residue mulching and potassium management options on growth and yield attributes of pearl millet [Pennisetum glaucum (L.) R. Br]. The experiment was conducted in a split plot design (SPD) with three mains plots, viz. C0, No residue mulching; C1, Crop residue mulching @3 t/ha; and C2, Crop residue mulching @3 t/ha + Pusa decomposer; and five potassium management in sub plot, viz. K0, No potassium application; K1, 40 kg K2O/ha; K2, 40 kg K2O/ha + potassium solubilising bacteria (KSB); K3, 30 kg K2O/ha + KSB; and K4, 20 kg K2O/ha + KSB. Pearl millet variety Pusa Composite 701 was used for the experiment. Significant variation among growth and yield parameters were observed under crop residue as well as potassium management options. Under crop residue mulching these were recorded maximum in treatment C2 (crop residue mulching @3 t/ha + Pusa decomposer). However, under potassium management options plant height (11.8–13.3%), ear head length (9.5 and 9.8%) and yield (26.7 and 24%) of pearl millet was recorded maximum in treatment K2 (40 kg K2O/ha + KSB) during the consecutive year over the control. These finding highlights the synergistic effects of potassium and residue management demonstrating that the presence of KSB enhanced potassium uptake by the plants, resulting in improved outcomes.

Genomic selection for agronomical phenotypes using genome-wide SNPs and SVs in pearl millet.

Pearl millet is an essential crop worldwide, with noteworthy resilience to abiotic stress, yet the advancement of its breeding remains constrained by the underutilization of molecular-assisted breeding techniques. In this study, we collected 1,455,924 single nucleotide polymorphism (SNP) and 124,532 structural variant (SV) markers primarily from a pearl millet inbred germplasm association panel consisting of 242 accessions including 120 observed phenotypes, mostly related to the yield. Our findings revealed that the SV markers had the capacity to capture genetic diversity not discerned by SNP markers. Furthermore, no correlation in heritability was observed between SNP and SV markers associated with the same phenotype. The assessment of the nine genomic prediction models revealed that SV markers performed better than SNP markers. When using the SV markers as the predictor variable, the genomic BLUP model achieved the best performance, while using the SNP markers, Bayesian methods outperformed the others. The integration of these models enabled the identification of eight candidate accessions with high genomic estimated breeding values (GEBV) across nine phenotypes using SNP markers. Four candidate accessions were identified with high GEBV across 22 phenotypes using SV markers. Notably, accession 'P23' emerged as a consistent candidate predicted based on both SNP and SV markers specifically for panicle number. These findings contribute valuable insights into the potential of utilizing both SNP and SV markers for genomic prediction in pearl millet breeding. Moreover, the identification of promising candidate accessions, such as 'P23', underscores the accelerated prospects of molecular breeding initiatives for enhancing pearl millet varieties.

Delineation of Selection Criterion Using Correlation and Path Analysis in Forage Pearl Millet (Pennisetum glaucum (L.) R. Br.) in North Gujarat Condition

Knowledge of genetic and phenotypic association among economic traits helps plant breeders in outlying efficient breeding strategies for development of high yielding forage pearl millet variety. An experiment was conducted at the research farm of Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat to evaluate plant characteristics associated with green forage yield and its attributes in forage pearl millet [Pennisetum glaucum (L.) R. Br.] genotypes in north Gujarat condition through correlation and path analyses during 2023. Twelve independent and 1 dependent variables were evaluated for the character association analysis of the 30 forage pearl millet genotypes. The evaluation was performed in a Randomized Complete Block Design with three replications. Data collection was done from 5 randomly selected plants and this data was used for analysis using R Studio. The green forage yield per plant had positive and significant correlation with plant height, number of tiller per plant, stem thickness, leaf length, leaf width, and dry fodder yield per plant at both genotypic and phenotypic level. Path analysis revealed the importance of leaf width and dry fodder yield per plant by showing high and positive direct effects towards green forage yield per plant. These characters also exhibited significant and positive association with green forage yield per plant at the genotypic and phenotypic. Hence, these traits could be considered as the vital component characters for development of high yielding genotypes in forage pearl millet.

Evaluation of Nitrogen Fertilizer Supply and Soil Nitrate Thresholds for High Yields of Foxtail Millet

Foxtail millet is an important cereal crop in the North China Plain. However, excessive nitrogen fertilizer application over the years has led to declining yield and soil quality. This study investigated nutrient management strategies for foxtail millet based on crop yield levels and soil nutrient availability. In a field where targeted fertilization was conducted over six seasons, nitrogen fertilization effects and the dynamics of soil-available nitrogen were monitored continuously for two consecutive years (2022–2023) across five different foxtail millet varieties with varying yield levels. The study aimed to determine the optimal nitrogen application rate for achieving a high yield of foxtail millet, the minimum soil nitrate threshold required to maintain soil fertility, and the effective nitrogen application rate range for sustaining soil-available nitrate levels. Results showed that fertilization significantly affected dry matter weight during flowering, while variety affected dry matter weight at maturity. The average nitrogen application rate for achieving high yield across all five millet varieties was 141.3 kg·ha−1. Specifically, the average nitrogen application rate of nitrogen-efficient varieties achieving high yield (5607.32–5637.19 kg·ha−1) was 151.5 kg·ha−1, while the average nitrogen application rate of nitrogen-inefficient varieties achieving high yield (4749.77–4847.74 kg·ha−1) was 134.5 kg·ha−1. Soil NH4+-N and NO3−-N content increased when nitrogen application rate exceeded 360 kg·ha−1, posing environmental risks. To achieve high yield, soil nitrate levels would be maintained at an average of 17.23 mg·kg−1 (before sowing) and 9.75 mg·kg−1 (at maturity). A relationship between soil nitrate and nitrogen application rate was established: y = 867.5 − 50z (where y represents the optimal nitrogen application rate for high yield (kg·ha−1), and z represents soil NO3−-N content in the 0–20 cm layer before sowing, ranging from 10.0 to 17.35 mg·kg−1), which provided a practical method for nitrogen fertilization to achieve high yield of foxtail millet. In this study, the fertilization strategy was optimized according to soil nutrient level and yield targets, and the nitrogen application rate was controlled within 360 kg·ha−1 based on the soil nitrate nitrogen content, which will be instructive for reducing fertilizer use, maximizing fertilizer efficiency, and increasing yield.

Evaluation of Released Finger Millet Varieties in the East Wollega Zone of Western Oromia for Yields and Acid Soil Tolerance

On the sub humid regions of western Oromia, many finger millet cultivars were released and produced, but their performance on acid soils was not well researched. A field experiment consisted of two factors (nine finger millet verities and two lime rates, limed and un-limed treatments) were laid out in factorial arrangement, Randomized Complete Block Design with three replications. The experiment was carried out at three locations (Bako, Diga and Gute) during 2019 and 2020 main cropping seasons to identify and recommend finger millet varieties better performing to acid soils of sub humid areas of western Oromia. The interaction effect of variety by lime for two years over three locations was significant for biomass weight and grain yield. Bako 09 produced significantly the highest mean biomass weight and produced increments of 10% biomass weight and 7% grain yield by application of lime compared to un-limed treatment. At Bako, Kumsa, Bako 09, Boneya and Addis 01 produced significantly the highest biomass weight and grain yield compared to others but not significantly different from each other and produced 4% to 12% yield advantage over all mean grain yield. At Diga and Gute, Bako 09 produced significantly high grain yield. Consequently, application of lime increased finger millet grain yield, soil pH, available P, organic carbon and total nitrogen. Considering higher grain yield, production of Bako 09 under limed condition is recommended for the sub-humid western Oromia. Besides lime application and varietal selection, cropping system managements like rotation, integrated use of organic and inorganic fertilizers, and split application of fine particle lime to acid soils require further research attention for sustainable crop production in acidic soils.

Effect of Nano Urea on Growth, Yield and Nutrient uptake of Finger Millet (Eleusine coracana L.)

Aim: The foliar application of nano urea is aimed at reducing the soil application of conventional urea. Nano fertilizers enhance crop growth and yield while enhancing nutrient uptake and reducing costs. They provide precise nutrient management, match crop growth stages, and offer an increased surface area for metabolic reactions. This boosts photosynthesis, leading to higher dry matter production and crop yield. Foliar application of nano urea (liquid) at tillering and booting stages in finger millet fulfills the nitrogen requirement and reflects higher crop productivity. Higher productivity of crops in a sustainable manner could be achieved by applying an appropriate combination of conventional fertilizers and nano fertilizers. Study Design: The experiment was laid out in split design with three replications. Place and Duration of Study: A field experiment was conducted in deep black soils at Agricultural Research Station, Siruguppa, during kharif 2022. Methodology: There were sixteen treatment combinations, consisting of different doses of nitrogen (no nitrogen, 50% RDN, 75% RDN and 100% RDN) with different concentrations of nano urea and conventional urea sprayed at tillering and booting stages for HR-13 variety of finger millet. Results: Application of 100% RDN has recorded significantly higher plant height, tillers plant-1, total dry matter accumulation, grain yield and straw yield (129.82 cm, 10.10, 36.17 g plant-1, 36.87 q ha-1 and 71.72 q ha-1, respectively) in comparison to no nitrogen and 50% RDN. However, 75% RDN (123.04 cm, 9.61, 34.02 g plant-1, 34.84 q ha-1 and 68.01 q ha-1, respectively) found on par with 100% RDN. Among foliar nitrogen, spray of nano urea @ 4 ml L-1 recorded significantly higher plant height (121.59 cm), tillers plant-1 (9.37), total dry matter accumulation (33.16 g plant-1), grain yield (31.97 q ha-1) and straw yield (65.52 q ha-1) compared to 0.5% and 1% conventional urea. However, nano urea @ 2 ml L-1 was on par with 4 ml L-1 nano urea and 1% conventional urea. Similar trend was recorded with regard to nutrient uptake by finger millet. Conclusion: Foliar application of nano urea reduced the dosage of conventional urea by 25 per cent.

Genome-wide assessment of population structure and association mapping for agronomic and grain nutritional traits in proso millet (Panicum miliaceum L.)

Proso millet is an important but under-researched and underutilized crop with the potential to become a future smart crop because of its climate-resilient features and high nutrient content. Assessing diversity and marker-trait associations are essential to support the genomics-assisted improvement of proso millet. This study aimed to assess the population structure and diversity of a proso millet diversity panel and identify marker-trait associations for agronomic and grain nutrient traits. In this study, genome-wide single nucleotide polymorphisms (SNPs) were identified by mapping raw genotyping-by-sequencing (GBS) data onto the proso millet genome, resulting in 5621 quality-filtered SNPs in 160 diverse accessions. The modified Roger's Distance assessment indicated an average distance of 0.268 among accessions, with the race miliaceum exhibiting the highest diversity and ovatum the lowest. Proso millet germplasm diversity was structured according to geographic centers of origin and domestication. Genome-wide association mapping identified 40 marker-trait associations (MTAs), including 34 MTAs for agronomic traits and 6 for grain nutrients; 20 of these MTAs were located within genes. Favourable alleles and phenotypic values were estimated for all MTAs. This study provides valuable insights into the population structure and diversity of proso millet, identified marker-trait associations, and reported favourable alleles and their phenotypic values for supporting genomics-assisted improvement efforts in proso millet.

Establishment of near-infrared rapid prediction model and comprehensive evaluation model for foxtail millet quality

Foxtail millet (Setaria italica) is one of the earliest cereal crops in the world and is still an important food crop in Asia, Europe and Africa. However, there are few reports on rapid and comprehensive evaluations of foxtail millet quality. In this study, color, nutrient content, cooking quality and sensory quality of 45 varieties of foxtail millet from the main production areas of the country were determined. Near infrared spectroscopy (NIRS) combined with partial least squares regression (PLSR) was developed for rapid and nondestructive detection of foxtail millet quality, where NIRS models for ash, amylopectin, protein, and VB1 and VB2 were desirable. In addition, correlation analysis, principal component analysis and cluster analysis were used for comprehensive evaluation of millet quality. The results showed that foxtail millet with low ash, protein, fat and fiber content and high amylopectin and carotenoid content has good cooking quality and was more preferred. This research will contribute to the quality evaluation and further processing of foxtail millet.

Finger Millet (<i>Eleusine Coracana</i> (L.) Gaertn) Production Status, Challenges, and Seed Source in Ethiopia; A Review

Similar to other cereal crops finger millet is being produced in a large volume in Ethiopia, as they are the principal staple food crop. Finger millet is considered as underutilized cereal crop which is commonly grown in the northern, west-northern, and western parts of the country and it is considered as a crop which is important to overcome malnutrition. Regions including Tigray, Amhara, Oromiya, Benishangul-Gumuz, Southern Nation and Nationalities Peoples (SNNP) and Gambela are considered as the potential regions for the production of finger millet in Ethiopia. Relatively, it is considered as drought tolerant as compared to other major cereals such as rice, wheat, and barley. In addition to this finger millet is considered as a highly resilient crop which can grow in different weather conditions, including drought, floods, and marginal soils. In Ethiopia it is commonly grown for the preparation of local foods like ‘injera’ and porridge and local drink such as ‘areki’ or ‘tella’. About 26 varieties of finger millet are cultivated in main season in the country. In Ethiopia, there are a limited source of cereal crop seeds which includes, farmer’s own saved seeds and government seed distributing companies. In this review the status of production, seed source and nutritional importance of finger millet in Ethiopia is presented.

Ancient Genome of Broomcorn Millet from Northwest China in Seventh Century CE: Shedding New Light to Its Origin and Dispersal Patterns

Broomcorn millet (Panicum miliaceum) is among the earliest domesticated staple crops in the world’s agricultural history and facilitated the development of several early agrarian cultures, particularly those originating in northern China. However, the propagation route of broomcorn millet in China from the Middle Ages to the present remains unclear. The aim of this study is to explore the genetic affinity between ancient and modern millet samples, trace the genetic origins and diffusion pathways of broomcorn millet, and provide insights into its domestication and spread. To achieve this, we sequence ancient DNA from broomcorn millet remains excavated from the Chashan Village cemetery (AD 691) in Gansu Province, China. Phylogenetic and population genetic analyses, integrating ancient and modern millet genomes, reveal a close genetic relationship between ancient millet and contemporary millet from Ningxia Province (445 km away from Chashan Village), suggesting a potential origin for the Chashan millet. This finding aligns with the tomb’s epitaph, which documents the reburial of the tomb’s owner, who was originally buried in Ningxia, and provides important archaeological evidence for understanding the interaction between geopolitical dynamics and the natural environment in northwest China during the late seventh century. Furthermore, outgroup-f3 and D statistics evidence suggests substantial genetic interactions between ancient millet and modern varieties from the Loess Plateau, Huang-Huai-Hai Plain, and Northeast Plain, indicating the dispersal route of broomcorn millet, along with human migration routes, from the northwest to northern China and ultimately to the northeast region, starting from the Middle Ages onward. This study enhances our understanding of millet’s genetic history, offers a novel perspective on burial archaeology, and provides valuable insights into the origins, domestication, and diffusion of broomcorn millet.

Transcriptome analysis and characteristics of drought resistance related genes in four varieties of foxtail millet [Setaria italica]

Foxtail millet [Setaria italica] plays a crucial role as a multigrain crop in agricultural production. However, due to future extreme weather conditions, drought remains the main abiotic stress that limits foxtail millet yield, it is highly significant to screen for drought-tolerant varieties throughout the entire growth period and identify the regulatory genes associated with drought resistance in foxtail millet breeding. We identified 217 foxtail millet seed resources for drought resistance during the maturity stage in the field, and subsequently categorized them into different levels of drought resistance. Two cultivars with extremely strong drought resistance during the maturity stage in the field, JKH4 (Chi 5422) and JKH6 (Chigu 26), as well as two cultivars with extremely weak drought resistance during the maturity stage in the field, JRK3 (17M1309) and JRK6 (Canggu 9), were selected for physiological comparison and transcriptome sequencing before and after drought treatment. Transcriptome analysis at the seedling stage revealed that JRK3 and JRK6 cultivar primarily regulated phenylpropanoid biosynthesis, MAPK signaling pathogen-plant, and plant hormone signal transduction pathway in response to drought stress. On the other hand, the fatty acid elongation pathway of JKH4 and JKH6 variety was found to be more significant. Furthermore, 22 drought resistance related genes were screened through transcriptome analysis of four foxtail millet varieties. These findings could offer valuable theoretical guidance for breeding foxtail millet with enhanced drought resistance and potentially facilitate the development of genetically engineered drought-resistant foxtail millet varieties.

Advanced physico-chemical and nutritional characterization of biofortified pearl millet varieties: A comparative evaluation of Dhanshakti and AHB-1200

Advanced physico-chemical and nutritional characterization of biofortified pearl millet varieties: A comparative evaluation of Dhanshakti and AHB-1200

Evaluation of Recently Released Finger Millet Varieties for Their Adaptability in West Haraghe Zone, Eastern Ethiopia

Finger millet is a major grain crop in the west hararghe zone. However, due to major constraints like lack of improved varieties and drought, the productivity is by far lower than the genetic potential of a crop in the study areas. Thus, current study initiated to obtain high-yielding and stable varieties. The study was conducted in districts of Habro, Mechara, and Gamachis of the west hararghe zone, using eight improved and one standard check finger millet varieties at 2020 main cropping seasons. The experiment was laid down in a randomized completely block design with three replications. Analysis of variance for grain yield across locations showed significant differences at p< 0.05. Further analysis of AMMI indicated that environments, varieties, and their interaction effects were significantly different. Even if, tested materials showed a significantly different grain yield across locations nevertheless, the GGE bi-plot analyses implied relatively high yielding and consistent across environments for varieties Bako-09, Gudetu, and Addis-01. Therefore, these varieties of finger millet were recommended for further evaluation at the farmer’s field.

Grain productivity and nutrient accumulation in millet straw grown in the off-season

Given the growing demand for grains cultivation in the off-season, an experiment was conducted to evaluate grain yield and nutrient accumulation in straw, straw production, and decomposition of pearl millet varieties and hybrids cultivated at different sowing times in off-season in the municipality of Rio Verde, state of Goiás. This was a randomized block experimental design, with four replications and four planting times. The work was carried out on an Oxisol, using two forage varieties, ADR300 and ADR500, the dual-purpose pearl millet hybrid ADR8010 for grain and biomass production, and two grain hybrids, ADR9010 and ADR 9020. The accumulation of macronutrients (nitrogen, phosphorus, potassium, calcium, magnesium and sulfur) was quantified in the straw at grain harvest. Hybrids ADR 9010 and ADR 9020 had the highest grain yield in the first season, with 2,626 and 2,425 kg ha-1, respectively, while the ADR 9010 hybrid showed the highest accumulation of macronutrients (except phosphorus) in the straw. Hybrid ADR 9010 is the best option for planting in the off-season with the objective of grain production, with high biomass production and accumulation of nutrients in the straw.

Insect Pest Management in Stored Millets

Recognizing the significance of millets as Nutri-Cereals, India celebrated the National Year of Millets in 2018, and subsequently, as proposed by India, the FAO of the United Nations declared 2023 as the International Year of Millets (IYoM). Consequently, there has been an intensive drive, globally, to increase the production and popularise the consumption of whole as well as processed millets by creating an awareness about their significant health benefits and richness in nutritional elements. Besides food, millets play a role as feed and in industrial uses. In commensurate with anticipated higher production, storage and international trade of whole millets and their products, it is important to protect millets throughout the supply chain from stored grain insects, the predominant biotic agent responsible for their qualitative and quantitative losses. In this context, this review focusses on the type of insect infestation encountered in millets from field to consumer levels affecting their market value and causing consumer concern. Insect management techniques adopted by the stakeholders such as farmers, traders, and government grain storage agencies are discussed. Due to financial constraints, small holder farmers use traditional storages and insect management measures using locally available plant products. Like other cereals, fumigation with aluminium phosphide (phosphine) preparations has a major role in insect pest management in millets at various storage levels. Hermetic storage bags of different capacities are used in India and in other millet producing countries especially at farmers level as a valuable insect management tool for whole millets and seed materials. Powdered and processed millets have shorter shelf-life and are more vulnerable to insect attack during storage. Hence, there are studies to increase their shelf life and to check infestation by suitable packaging material. Alternate insect management techniques investigated include ozone treatment, protective packaging materials, screening and breeding of millet varieties that are resistant to grain insects National regulations pertinent to insect management in stored millets have also been discussed.

Demonstration by New Finger Millet Variety ATL 1 to Improve the Productivity in Villupuram District of Tamil Nadu, India

Demonstration by New Finger Millet Variety ATL 1 to Improve the Productivity in Villupuram District of Tamil Nadu, India

Genome-Wide Identification, Bioinformatic Characterization, and Expression Profiling of Starch Synthase (SS) Genes in Foxtail Millet under Drought Condition

Millet, a vital and nutritionally dense cereal extensively cultivated in Sub-Saharan Africa, plays a key role in ensuring food security. This study investigates the starch synthase (SS) gene family, which is crucial for starch biosynthesis and influences various plant functions and stress responses. While the specific roles of SS genes in millet under drought conditions are not fully elucidated, this research provides a thorough analysis of the SS gene family in millet. A total of twelve millet SS genes (SiSSs) were identified and classified into four subfamilies (I–IV) through gene structure and phylogenetic analysis. The SiSS genes were unevenly distributed across millet chromosomes, with cis-acting elements associated with plant growth and stress defense being identified. Quantitative PCR (qPCR) revealed dynamic and varied expression patterns of SiSSs in different tissues under drought stress. Millet plants subjected to drought conditions showed higher tissue starch content and increased starch synthase activity compared to controls. Importantly, the expression levels of the twelve SiSSs were positively correlated with both starch content and synthase activity, suggesting their significant role in drought tolerance. This study enhances our understanding of the millet SS gene family and highlights the potential of these genes in breeding programs aimed at developing drought-resistant millet varieties. Further research is recommended to validate these findings and delve deeper into the mechanisms underlying drought tolerance.

Performance of Finger Millet Varieties (Eleusine coracana L.) in Different Establishment Methods

Performance of Finger Millet Varieties (Eleusine coracana L.) in Different Establishment Methods