Genetic Effects of Ultra-High-Diluted-Biomedicines Gall 30C, Gall 200C, and Gall 1000C may be Vaccines against Plant and COVID-19 Diseases: Improved Agriculture-Health-Medical-Pharmaceutical-Science-Technology-Communication-Issues!

The current coronavirus 2 (SARS-CoV-2) outbreaks of Coronavirus disease 2019 (COVID-19) pandemic have emphasized the vulnerability of human populations to novel viral pressures, causing an emergent global pandemic and badly impacts on agriculture-environment-health-socio-economy medical-pharmaceutical-science-technology-communication-issues. And India emphasis on okra, Abelmoschus esculentus (L.) Moench Cv. Ankur-40, the ‘Nature’s-Gift to Human-Disease-Free-Healthy-Life’ and the most ‘Economically-Important Number-One Consumption-Vegetable-Crops,’ is damaged by different pathogens like nematodes, causing the root-knot (RK) disease which is easily controlled by different chemical-pesticides, and so it is an urgent need to develop potential epidemiological and biomedical vaccines. The Ultra-High-Diluted Biomedicines (UHDBM) Gall 30C, Gall 200C, and Gall 100C, prepared from Okra Root-Galls (ORG) or Gall MT, applied by foliar spray @ 20 mL/plant each group respectively, are highly effective against the root-knot disease of okra, with increasing fresh-plant growth and fruit production. The UHDBM; Gall 30C, Gall 200C, and Gall 1000-pretreatments are more effective than the untreated ones, and Gall 200C shows the most potential result in all respects. The genetic-effects of UHDBM thought to induce systemic acquired resistance response of the treated plants through the expression of pathogenesis-related (PR)proteins-genes (22 to 4 numbers), which are more or less similar molecular range (295 kD to 11 kD) of many coronaviruses, and it will responsible for preventing RK and COVID-19 like variant-virus diseases by inducing defense-resistance or increasing innate-immunity, and enriching agricultureenvironment-health-socio-economy-medical-pharmaceutical-science-technology-communication-issues (AEHSEMPSTCI) with the toxic-free world, and it may help to develop best potential new preventive treatments methods or drug or vaccines, in the field of ‘21st Century COVID-19 like Pandemic in the new normal situation in future.

India emphasis on okra (lady finger) for the 'food security' which is the most important and crucial aspect of sustainable development in the agricultural sector and the backbone of the economy, but badly impact of the COVID-19 pandemic on food security, agriculture, massive consequences on health and livelihoods of developing countries [21], and it not only reduced incomes, but also disrupted supply chains, chronic and acute hunger was on the rise due to various factors including conflict, socio-economic conditions, natural hazards, climate change, and pests like the locust outbreak compounding this crisis across 23 countries with the other zoonotic diseases remain a recurrent threat. And it is reported that the okra, Abelmoschus esculentus (L.) Moench, is one the most economically important, commercially exploited, number one consumption in a variety of ways vegetable crops, which is the oldest widely cultivated oligo purpose, significantly contribute for nutritional, medicinal and industrial application , and used as traditional medicine, achieving the 'Nature's Gift to human disease-free healthy life' multipurpose crop, achieving India first in the world for the fruits rich in vitamins, calcium, folic acid, carbohydrates, phosphorus, magnesium and potassium, iodine, mineral matters, and a good source of superior nutritional quality for human nutrition for preventing different diseases; like cardiovascular disease, type 2 diabetes, kidney diseases, skin infection, digestive diseases, some cancers, antioxidant, nootropic, eye, body immunity, blood pressure, obesity, asthma, constipation, heart disease, sexual health, and neurological disorders, etc., and mature fruit and stems contain crude fiber, used in the paper industry and sugarcane industry [22][23][24][25]. The enrich source of nutrients, minerals, and fibers of okra has invited to many pathogens, pests, and diseases infestation, and only the nematode pathogens causing root-knot, damage 10-40% of the total crop production annually, causing serious problems in our country and this indirectly affects our advanced agronomy-plantbreeding-horticulture-environment socio-economy green-sciencetechnology-communication-issues, and agricultural-economy also. Though the pesticides are very much effective but it creates several problems in toxicity, pollution, cost-effectiveness, environmentfriendliness, and biodiversity conservation. On the other hand the pesticides block functional first known gene transfer between plants to insects or animals, which is used in the host's defenses mechanism for new pest-control strategies also, and recently in the 31countries of the globe that higher airborne pollen concentrations correlated with increased SARS-CoV-2 infection rates .
Recently after long lockdowns in Purba Bardhaman, West Bengal, India (Figure 1), only two day shows that the total COVID-19 positive cases are 37494-37524, the total number of discharge cases are 36685-36718, the total number of COVID-19 death is 436-436, rate of recovery is 97.84%-97.85% and rate of mortality is 1.16%-1.16% respectively. So it is an urgent need to find out by developing policyinitiative, cheap, non-phytotoxic, and non-pollutant potential highdiluted-biomedicines (UHDBM) for preventing both the pandemic crisis by improving the agriculture system with the findings and other new research to develop future support and treatments.

Aims and Objectives
The purpose of the current study is to use and confirm the systematic signaling and induced natural defense or immunity in the host plants, Abelmoschus esculentus (L.) Moench Cv. Ankur-40, by applying UHDBM; Gall 30C, Gall 200C, and Gall 100C, prepared from Gall MT (GMT) or Gall Roots (GR) as a preventive measure against root-knot, and COVID-19 diseases ideas, and also to find out the actual reasons of the genetic effects on the UHDBM for preventing diseases.
The current paper contributes the development of the most policy-initiative, cheap, non-phytotoxic, non-pollutant, and sideeffect free potential UHDBM global vaccines; Gall 30C, Gall 200C, and Gall 1000C, for controlling both the pandemic crisis; root-knot diseases, and the future-pandemic COVID-19 like virus diseases, by enriching agriculture-environment-health-socio-economymedical-pharmaceutical-science-technology-communication-issues (AEHSEMPSTCI) with the findings and other new research to drug development for the future support and the best treatments or potential vaccine for all.

Treatments with UHDBM Gall 30, Gall 200C, and Gall 1000C test solutions
The UHDBM; Gall 30C, Gall 200C, and Gall 1000C -test solutions, were applied into the okra plants and sprayed on plants @20 mL/ treated plants three days before inoculation for pretreatments with nematodes-J 2 respectively. Control okra plants were treated with an equal amount (20 ml) of control solutions prepared with sterile distilled water, and the plants were regularly watered in the morning and evening. During spraying, the soil surface underneath each plant was covered with a polythene sheet. Plants in both uninoculated untreated and inoculated untreated groups received a spray of an equal amount of control solutions. All treatments were done in hygienic conditions [3][4][5][6][7][8][9][12][13][14][15][16][17][18][19][26][27][28][29][30][31]. All the data were analyzed by ANOVA (Analysis of Variance). The experiment was repeated thrice. Data from the last experiment are reported here.
Harvesting: All the okra plants were uprooted 53 days after the sowing of seeds. The following measure measurement was taken: biomass of shoot, root, and fruits, root gall number, nematode population in roots (2 g) and soil (200 g), the protein content of root and fruits. Proteins were estimated by the Folin-phenol method [4,[14][15][16][17][18][19][28][29][30][30][31][32][33][34]. All the data were analyzed by Analysis of Variance (ANOVA). The experiment was repeated five times with similar results and the data from the third experiment were represented in table 2.

Effect on RK with UHDBM Gall 30, Gall 200C, and Gall 1000C
The pretreated with UHDBM; Gall 30, Gall 200C, and Gall 1000C significantly (P ≤ 0.05 by ANOVA) increased plant growth in terms of the fresh biomass of shoot and fruits compared to the inoculated and untreated plants ( Table 2). Root galls, nematode population in the root and soil, and root protein content were significantly (P ≤ 0.05 by ANOVA) reduced in pretreated plants as compared to the untreated ones ( Table 2). The number of fruits and protein content in green fruit was significantly reduced in inoculated untreated plants as compared to the uninoculated ones, and all the pretreatments with biomedicines; Gall 30, Gall 200C, and Gall 1000C showed better plant growth and lesser intensity of the root-knot disease, and Gall 200C showed the best results in all respect ( Table 2).  (Table 1 and Figure 2). The highest molecular weight of the OR-protein gene is 295 kD and the lowest molecular weight of the ORprotein genes is 11 kD. The lowest number of the new pathogenesisrelated protein genes (PR-proteins) is 4 in the uninoculated untreated okra roots, and the highest number of the new PR-proteins-genes is 22 in the HDBM-Gall 200C-pretreated okra-pretreatment-group, 18 in the Gall 30C, and the same number i.e. 14 number of PRproteins-genes is in both the ultra-high-diluted biomedicines Gall 1000C-pretreatment-and inoculated untreated -treatment-group respectively (Table 1 and Figure 2).
And the NF contained 18-proteins-genes and the molecular weight of the NF proteins-genes ranging from the lowest 12 kD to the highest 280 kD, and the total number of the PR-proteins-genes of NF is 16 in comparison to the uninoculated untreated group (Table 1 and Figure 2).

On growth
The present study clearly showed that the OP-growth in terms of fresh biomass of shoot and fruits, and the number of fruits was higher than inoculated untreated groups in all the pretreatment groups of OP treated with the UHDBM; Gall 30, Gall 200C, and Gall 1000C, but reverse in the fresh biomass of roots of inoculated untreated one [4,[14][15][16][17][18][19][28][29][30].

On RK diseases
All the pretreatment groups of OP treated with UHDBM; Gall 30, Gall 200C, and Gall 1000C, decreased RK-diseases in terms of nematode infection in root gall number, and nematode population in root in comparison to inoculated untreated groups, and the population of nematode in the rhizospheric soil was the maximum with the group treated with Gall 30, Gall 200C, and Gall 1000C-biomedicines, and minimum with the inoculated untreated group, due to potential UHDBM, and it is also showed that the UHDBM; Gall 30, Gall 200C, and Gall 1000C, might induce synthesis of some antagonistic substance in the treated-OP, which is proved from the inoculated untreated ORG contained the highest protein-content due to presence of a large number of nematodes [4,[14][15][16][17][18][19][28][29][30].

On toxicity
In the recent study is also showed that the UHDBM; Gall 30, Gall 200C, and Gall 1000C, had no direct toxic effects on nematodes juveniles, and OP, but it induced synthesis of some resistance substances in okra plants to M. incognita infection for preventing RKdiseases in the OP, and for these reasons, all the pretreated group had significantly greater fresh biomass of shoot-and fruits-plants than inoculated untreated one [4,[14][15][16][17][18][19][28][29][30].

On pathogenesis-related (PR) of root proteins-genes
It is known that the M. incognita is known to share common antigens with its host plants [48], and Iqbal S, et al. (2020) informed that the attempt to 'Silence Genes' of the root-knot nematode, M. incognita results in diverse responses including an increase and no change in expression of some genes. So in the all the treated plant's roots show that the UHDBM; Gall 30, Gall 200C, and Gall 1000C-pretreatments resulted in an increased number of proteins-genes in the root than inoculated untreated-okra plants-group; the highest number of PR-proteins-genes in the UHDBM; Gall 30, Gall 200C, and Gall 1000C-pretreated group is 23, and the next highest number of the PRprotein-gene is 19 in the pretreated-Gall 30C -inoculated group, 16 in the Gall 1000C group, and 15 in the inoculated untreated group, and the lowest number of protein is 11 in the uninoculated untreated group respectively, which proved and confirmed that during infection with the nematode, host plants showed minimal defense responses to the nematode because of this antigenic similarity, and the different PRproteins-genes of the ORG proteins ranging from 295 kD (the highest molecular weight protein) to 11 kD (the lowest molecular weight protein) of the OR-protein. And all the pretreated UHDBM; Gall 30, Gall 200C, and Gall 1000C-stimulate the synthesis of the number of different PR-proteins-antigens-genes that must induce defense responses in which the nematodes fail to survive [4,[14][15][16][17][18][19][28][29][30], and it is also proved from the plant-nematode interaction, newly synthesized PR-proteins genes have been found in potato plants infected with the potato-cyst-nematodes Globodera pallida and G. rostochiensis [4,7,18,19,24,25,28,30,46,47]. It is also reported that Salicylic Acid (SA) increases resistance in plants against RK-diseases by inducing expression and accumulation of pathogenesis-related-I protein (14 kD, PR-I) in the sprayed plant-root and leaves, and it sprays enhances-PAL higher activity in infected-roots [4,7,18,19,24,25,28,[39][40][41][42][43][44][45][46][47].

On PR-proteins-genes of NF
The sixteen PR-proteins-genes out of total eighteen proteins-genes of NF of M. incognita in comparison to the uninoculated untreated group, and the molecular weight of the nematode-proteins-genes ranging from 12 kD to 280 kD, already confirmed the potential efficacy of the UHDBM-Gall MT or GMT use as an effective stimulus for the expression of these various new 16 defense-related PRproteins-genes might be provided resistance to nematode-infection in okra plants due to nematodes present in the gall roots also, and it can be preventing pathogenesis in patients with COVID-19 due to more or less proteins-genes range (240 kD to 26 kD) in different bovine-and human-coronavirus-structural-proteins which send genetic-information, and the SARS-CoV-2 genes may integrate with human-DNA to code the essential nonstructural-proteins like an RNA-polymerase also, and the nematode-proteins-genes slip into human-chromosomes and the diverse immunological-factors on viral-dissemination, immunotherapeutic-options, and inflammatoryresponses, and need molecular-characterization and understanding of the human-coronavirus-life-cycle, structural and functional properties of SARS-CoV-2 spike-protein for potential antivirus-drug development, and analysis of therapeutic-targets for SARS-CoV-2 and discovery of potential-drugs by computational-methods, and genomic-characterization and epidemiology of 2019 or genomicepidemiology has come of age during this pandemic affording to track SARS-CoV-2 sequences helped identify worrying-variants, with implications for virus-origins and receptor-binding also, for preventing the COVID-19, but researchers are blind to emergingmutations in some-regions, and so it is thought, "the next pandemic by transforming food-systems for affordable healthy-diets" [39][40][41][42][43][44]. Recently it may help from the report that cellular senescence (SnC) contributes to inflammation, multiple-chronic-diseases, and age-related dysfunction, and the SnC become hyper-inflammatory in response to pathogen-associated-molecular-patterns (PAMPs), including SARS-CoV-2 Spike-protein-1, increasing expression of viral entry proteins and reducing anti-viral gene-expression in non-SnCs through a paracrine-mechanism [1,2,14-21].

On genetic effects of ORG-pathogenesis-related (PR)proteins-genes
In the UHDBM-Gall 200C -pretreated Okra Root Galls (ORG), the 22 PR-proteins-genes out of total 23 proteins-genes in comparison to the uninoculated untreated okra plant group, and the molecular weight of the ORGs-proteins-genes ranging from lowest 12.5 kD to highest 295 kD, proved the potential efficacy of the pretreatment-UHDBM -Gall 200C act as the most effective stimulus for the expression of these many new 22 defense-related PR-proteins-genes which might be provided resistance to nematode-infection in okra plant. All the UHDBM; Gall 30, Gall 200C, and Gall 1000C-pretreatments collectively resulted in an increased number of PR-proteins-genes. And the genetic effects of UHDBM; Gall 30, Gall 200C, and Gall 1000C-pretreatments resulted in an increased number of proteinsgenes, are thought to induce systemic acquired resistance response of all the pretreated plants through the expression of Pathogenesis-Related (PR)-proteins-genes (22 to 14 numbers), which are more or less similar molecular range (295 kD to 11 kD) of various coronavirus, and it will responsible for preventing RK and COVID-19 like virus diseases by inducing resistance of plants or increasing immunity of animals respectively, and enriched AEHSEMPSTCI [14][15][16][17][18][19][20][21].

Idea for Future Research
The UHDBM; Gall30C, Gall 200C, and Gall 1000C-pretreated ORG-proteins could be induced the production of new defense-related PR-genes in the test plants and might be confirmed, and in near future, synthetic production of the RG-proteins would be the most potential cost-effective personalized-biomedicine OR social vaccine OR vaccine against coronavirus-2 like pandemic diseases by increasing immunity, and helping policy initiative clinical research in all areas in the field of advanced agronomy-plant-breeding-horticulture-agricultural, aquatic sciences, environment, socio-economy, and green-science-technologycommunication issues by preventing okra root-knot and COVID-19 also. Thus, RG-proteins will serve as very effective biomedicines that would be the most effective cheapest, non-phytotoxic, non-pollutant, conserve our biodiversity, and this vaccine might be the most effective against the delta variant of coronavirus, and scholars that give good scope for new development and future research in various fields of pathology-medical-pharmacology preventing fungal infections by increasing the post-COVID weakened lungs as well as the immune also [14][15][16][17][18][19][20][21]. And in the near future UHDBM; Gall30C, Gall 200C, and Gall 1000C pretreatment GR may be used as mixing vaccines to provoke potent immune responses for the real-world efficacy and rare side effects. And also help for the future mitigation strategies, and COVID appropriate and risk behavior at slums different areas also [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19]24,[23][24][25][26][27][28][29][30].

Journal of Drug Research and Development
Open Access Journal

Conclusions
The UHDBM Gall 30C, Gall 200C, and Gall 1000C are again highly effective against the root-knot disease of okra, with increasing plant growth and fruit production. The UHDBM; Gall 30C, Gall 200C, and Gall 1000-pretreatments are more effective than the untreated ones, and Gall 200C shows the most potential result in all respects. The genetic-effects of UHDBM thought to induce systemic acquired resistance response of the treated plants through the expression of PR-proteins-genes (22 to 4 numbers), which are more or less similar molecular range (295 kD to 11 kD) of many coronaviruses, and it will be responsible for preventing plant and COVID-19 like variant-virus diseases by inducing defense-resistance or increasing innate-immunity, and enriching green-AEHSEMPSTCI with the toxic-free world, and it may help to develop best potential new preventive treatments methods or drug or vaccines, in the field of '21 st -Century COVID-19 like a pandemic in the new normal situation in future. And in the near future pretreatment of gall root may be used as mixing vaccines to provoke potent immune responses for the real-world efficacy and rare side effects, and it also may help for the future mitigation strategies, and COVID appropriate and risk behavior at slums different areas.