Progress 04/01/24 to 03/31/25
Outputs
Target Audience:Broader plant science community in general and cotton breeders and geneticists, in particular, will benefit from the research conducted under this project. The information obtained from the results and the technology developed under this project will enable breeders to create drought tolerant varieties. Thus far, one particular technology to emerge from this project was the refinement of grafting procedure in cotton. Published information regarding the grafting technique using in vitro-grown seedlings will be useful not just for cotton researchers, but also for scientists that are working with other important dicot crops. The method can be used by the scientists working with important dicot crops (e.g. cotton and soybean) to conduct basic research that includes investigating communication between shoot and root under normal conditions and also in response to various stresses, as well as understanding long-distance movement of proteins, RNAs, hormones, and other biochemicals. Changes/Problems:One problem we faced was the availability of sufficient quantity of seeds from most of the genotypes that have been shown to be either drought tolerant or drought sensitive,based on the published literature. Therefore, we are havingto grow these in greenhouses to obtain sufficient quantity of seeds to conduct grafting studies. A few exotic genotypes (Tex1236, Tex169) turned out to be photoperiod sensitive (short day) and did not flower in the greenhouse available to us at the University because of the supplementary lights from the neighboring greenhouse and proximity to streetlights. We had to shift these plants to a different greenhouse (local USDA-ARS Facility)at the start of winter (2023-2024) that are not exposed to streetlight or supplemental lighting from the neighboring greenhouses which can interfere with the flowering of these particular plants. Progenies from selected exotic parents is again being grown in the same greenhouse for seed multiplication during the current winter season (2024-2025).Some other commercial varieties that were deemed drought tolerant or drought sensitive (based on published results from field trials) were first examined for their tolerance to water deficit stress under growth chamber/greenhouse conditions. However, our initial tests did not provide clear results on their level of tolerance to drought stress. We have modified our experimental procedures to ascertain the response of various genotypes to water deficit stress. Once we have obtained sufficient quantities of seeds from various genotypes and established conditions to grow the plants approximating field environment, we will be able to conduct grafting experiments as outlined in our proposal. What opportunities for training and professional development has the project provided?A postdoctoral Research Associate (Dr. Shiva Ram Bhandari) has been working full-time on this project. Although his previous research experience was limited to biochemical analysis, he is being trained in physiological and molecular responses to water-deficit stress under this project. Participation in biweekly lab meetings is helping him to broaden his horizons in areas related to cell biology/genetic engineering/gene editing. He is also gaining experience in writing and presenting reports. How have the results been disseminated to communities of interest?A paper detailing the refinement of the grafting procedure has been published [Pandeya, D., Campbell, L.M. and Rathore, K.S. (2023) A Grafting Procedure for Gossypium hirsutum L. Based on In Vitro Grown Seedlings. Journal of Cotton Science 27: 149-157]. An oral presentation was given by the P. I. based on this work at the 2024 Beltwide Cotton Conferences [Pandeya, D., Campbell, L.M. and Rathore, K.S. (2024) Development of an efficient intervarietal grafting method for Gossypium hirsutum and it's use for interspecific grafting. 2024 Beltwide Cotton Conferences, Fort Worth, TX]. Another oral presentation was given by the P. I. on the use of grafting to study transport of biochemicals between shoot and root at the 2025 Beltwide Cotton Conferences [Rathore, K.S., Pandeya, D., Campbell, L.M., Puckhaber, L. and Suh, C. (2025) Lysigenous Glands in the Aboveground Parts of a Cotton Plant do not Depend on the Roots as a Source of Gossypol. 2025 Beltwide Cotton Conferences, New Orleans, LA]. What do you plan to do during the next reporting period to accomplish the goals?Now that we have identified some drought tolerant and drought sensitive genotypes, as indicated earlier, we will utilize these to generate reciprocal grafts that will then be subjected to drought stress. These grafted unions will then be evaluated for variousmorphological, physiological, biochemical and molecular parameters to understand the role of roots versusshoot in conferring tolerance to water deficit stress. We expect that theresults from these studies will demonstrate the utility of grafting as a tool to prescreen the cotton genotypes/species that have root systems with the ability to protect the shoot function and productivity under drought stress.Based on the results obtained in the proposed study, cotton breeders can use a similar approach to identify genetic markers for root characteristics and choose appropriate germplasm to create drought tolerant varieties while retaining superior fiber/seed yield and quality characteristics.
Impacts
What was accomplished under these goals?
We further refined the grafting technique that is based on in vitro-grown Gossypium hirsutum seedlings and showed that if hypocotyls of the seedlings providing the rootstocks and scions are similar in girth, it is possible to achieve 90-100% success in grafting. Furthermore, we were able to obtain intervarietal and interspecific grafting in cotton. The results were published in the Journal of Cotton Science (Pandeya et al., 2023). Two oral presentations have been given at the Beltwide Meetings (Fort Worth, TX, January 2024; New Orleans, LA, January 2025) on the refinement of grafting technique in cotton and its utility in studying translocation of biochemicals (gossypol and related terpenoids) between shoot and root. We relied on previously published studies or personal communications with scientists studying drought tolerance/susceptibility in cotton to obtain seeds and to screen and select genotypes that can be used to conduct our research pertaining to the objectives of our project. Some morphological (Root/shoot weight/length, leaf relative water content), physiological (stomatal conductance, photosynthesis and transpiration) and biochemical (proline, melondialdehyde and chlorophyll content) parameters are being used to assess the degree of tolerance to water deficit stress. Commercial cotton genotypes determined to be drought tolerant/susceptible based on two of the published studies (Aime et al., 2021; Wedegaertner et al., 2023) did not respond as expected in our growth room-based trials. Some of the exotic germplasm obtained from USDA Cottonseed Bank (Tex1236, Tex169, studied by Quisenberry and coworkers in 1980s) and few other genotypes (UGA230, CS50, Tipo Chaco and DP16 obtained from Duke Pauli, University of Arizona) were insufficient in quantity to conduct tolerance tests and to directly use in grafting experiments. Therefore, these are being grown in greenhouses for seed increase. The exotic genotypes (Tex1236, Tex169) turned out to be photoperiod sensitive (short-day). These were grown in greenhouses over the wintertime (2023/2024). Unfortunately, the flower color and other morphological features indicated that the seeds obtained from the USDA Cottonseed Bank were not pure, they were segregating. The progeny obtained from the two lines was tested in a growth room setting to assess their relative abilities to tolerate water-deficit stress. Results from small-scale experiments with 'selected' progeny of Tex1236, Tex169 appears to be in line with the findings of Quisenberry et al. Based on the results, one plant from each of the two germplasms (Tex1236, Tex169) was chosen for the project. The amount of seeds obtained from these two selected plants, however, were low, therefore, these were planted and are being grown again this winter (2024/2025) to obtain more seeds. This material will be used to conduct the grafting studies to dissect out the roles of root versus shoot as outlined in our proposal.Based on their wilting score and recovery following drought stress and certain physiological parameters, it seems that CS50 is more tolerant to drought stress whereas Tipo Chaco is less tolerant. DP-16 also appears to show slightly better tolerance to water-deficit stress. Again, the supply of seeds obtained from our collaborator was very low and therefore, we had to multiply the seeds before conducting the experiments. Seed multiplication, harvesting and ginning for these three genotypes is still ongoing. In addition to these genotypes, we have recently obtained some seeds of the following genotypes: DP14, Station Miller, DES56, Paymaster HS26 (tolerant) and Toole, Paymaster HS200 (sensitive) based on a study conducted by one of our collaborators (Ghimire et al., 2024). These are being grown in a greenhouse for seed multiplication. Now that we have identified some genotypes that exhibit either tolerance or susceptibility to drought stress, we will utilize these to conduct the studies based on intergenotypic grafting to understand the contribution of root versus shoot in conferring drought tolerance in cotton.
Publications
- Type:
Peer Reviewed Journal Articles
Status:
Published
Year Published:
2023
Citation:
Pandeya, D., Campbell, L. M. & Rathore, K. S. (2023) A Grafting Procedure for Gossypium hirsutum L. Based on In Vitro Grown Seedlings. J. Cotton Sci. 27: 149-157.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
RATHORE, K. S., PANDEYA, D. & CAMPBELL, L. M. (2024) Development of an Efficient Intervarietal Grafting Method for Gossypium hirsutum and its Use for Interspecific Grafting. 2024 Beltwide Cotton Conferences. Fort Worth, TX.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2025
Citation:
Rathore, K.S., Pandeya, D., Campbell, L.M., Puckhaber, L. and Suh, C. (2025) Lysigenous Glands in the Aboveground Parts of a Cotton Plant do not Depend on the Roots as a Source of Gossypol. 2025 Beltwide Cotton Conferences, New Orleans, LA
|
Progress 04/01/23 to 03/31/24
Outputs
Target Audience:Broader plant science community in general and cotton breeders and geneticists, in particular, will be able to utilize the tools developed under this project to create drought tolerant varieties. Thus far, one particular technology to emerge from our work was the refinement of grafting procedure in cotton. This method can be used by the cotton community to conduct basic research that includes investigating communication between shoot and root under normal conditions and also in response to various stresses, long-distance movement of proteins, RNAs, hormones, and other biochemicals. Changes/Problems:One problem we face is the availability of sufficient quantity of seeds from most of the genotypes that have been shown to be either drought tolerant or drought sensitivebased on the published literature. Therefore, we are havingto grow these in greenhouses to obtain sufficient quantity of seeds. A few exotic genotypes turned out to be photoperiod sensitive (short day) and did not flower in the greenhouse available to us at the University because of the supplementary lights from the neighboring greenhouse and proximity to streetlights. We had to shift these plants to a different greenhouse (local USDA-ARS Facility)at the start of winter that are not exposed to streetlight or supplemental lighting from the neighboring greenhouses which can interfere with the flowering of these particular plants.Some other commercial varieties that were deemed drought tolerant or drought sensitive (based on published results from field trials) were first examined for their tolerance to water deficit stress under growth chamber/greenhouse conditions. However, our initial tests did not provide very clear results on their level of tolerance to drought stress. We are now modifying our experimental procedures to ascertain their response to water deficit stress. This involves use of long (18" long), tubular DeepotsTM, instead of the small, square pots and also different brand of commercial soil medium. We believe that once we have obtained sufficient quantities of seeds from various genotypes and established conditions to grow the plants approximating field environment, we will be able to conduct experiments as outlined in our proposal. What opportunities for training and professional development has the project provided?A F/T postdoctoral research associate and a P/T technician have been trained and working on this project. How have the results been disseminated to communities of interest?A paper detailing the grafting procedure has been published [Pandeya, D., Campbell, L.M. and Rathore, K.S. (2023) A Grafting Procedure for Gossypium hirsutum L. Based on In Vitro Grown Seedlings. Journal of Cotton Science 27: 149-157]. Also, an oral presentation was given by the P. I. based on this work at the 2024 Beltwide Cotton Conferences [Pandeya, D., Campbell, L.M. and Rathore, K.S. (2024) Development of an efficient intervarietal grafting method for Gossypium hirsutum and it's use for interspecific grafting. 2024 Beltwide Cotton Conferences, Fort Worth, TX]. What do you plan to do during the next reporting period to accomplish the goals?Once we have identified one or more drought tolerant and drought sensitive genotypes, indicated earlier, we will utilize these to generate reciprocal grafts that will then be subjected to drought stress. These grafted unions will then be evaluated for variousmorphological, physiological, biochemical and molecular parameters to understand the role of roots andshoot in conferring tolerance to water deficit stress. We expect that theresults from these studies will demonstrate the utility of grafting as a tool to prescreen the cotton genotypes/species that have root systems with the ability to protect the shoot function and productivity under drought stress.Based on the results obtained in the proposed study, cotton breeders can use a similar approach to identify genetic markers for root characteristics and choose appropriate germplasm to create drought tolerant varieties while retaining superior fiber/seed yield and quality characteristics.
Impacts
What was accomplished under these goals?
We further refined the grafting technique that is based on in vitro-grown Gossypium hirsutum seedlings and showed that if hypocotyls of the seedlings providing the rootstocks and scions are similar in diameter, it is possible to achieve 90-100% success in grafting. Further, we were able to obtain intervarietal and interspecific grafting in cotton. The results were published (Pandeya et al., 2023). Based on the results from published studies and personal communications, we have obtained several seeds of cotton genotypes that are potentially either drought tolerant (G. hirsutum: PHY499 WRF, PHY400 W3FE, Tex1236, UGA230, CS50;G. barbadense: GB259 and Gb189) or drought sensitive (NG5007 B2XF, DP2115 B3XF, Tex169, Tipo Chaco, DP16). Some of the exotic germplasm obtained from USDA Cottonseed Bank (Tex1236, Tex169, GB259 and Gb189) and few other genotypes (UGA230, CS50, Tipo Chaco and DP16) were insufficient in quantity to directly use in grafting experiments. Therefore, we are growing these in greenhouses for seed increase. The exotic genotypes turned out to be photoperiod sensitive (short-day). These were grown in greenhouses over the wintertime and are just beginning to produce seeds. Once we obtain sufficient quantities of seeds, we will perform stress tolerance experiments with the grafts. Some other commercial varieties that were deemed drought tolerant or drought sensitive (based on published results from field trials) were first examined for their tolerance to water deficit stress under growth chamber/greenhouse conditions. Some morphological (Root/shoot weight/length, leaf relative water content), physiological (stomatal conductance, photosynthesis and transpiration) and biochemical (proline, melondialdehyde and chlorophyll content) parameters were used to assess the degree of tolerance to water deficit stress. However, our initial tests did not provide very clear results on their ability to tolerate drought stress. We are now modifying our experimental procedures to ascertain their response to water deficit stress.?
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Pandeya, D., Campbell, L.M. and Rathore, K.S. (2023) A Grafting Procedure for Gossypium hirsutum L. Based on In Vitro Grown Seedlings. Journal of Cotton Science 27: 149-157.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Pandeya, D., Campbell, L.M. and Rathore, K.S. (2024) Development of an efficient intervarietal grafting method for Gossypium hirsutum and it's use for interspecific grafting. 2024 Beltwide Cotton Conferences, Fort Worth, TX
|