|
xml version 1.0 encoding UTF-8 standalone no
fcla fda yes
!-- Advancement of Cowpea Into a Dual Edible and Ornamental Crop ( Book ) --
METS:mets OBJID AA00092429_00001
xmlns:METS http:www.loc.govMETS
xmlns:xlink http:www.w3.org1999xlink
xmlns:xsi http:www.w3.org2001XMLSchema-instance
xmlns:daitss http:www.fcla.edudlsmddaitss
xmlns:mods http:www.loc.govmodsv3
xmlns:sobekcm http:digital.uflib.ufl.edumetadatasobekcm
xmlns:lom http:digital.uflib.ufl.edumetadatasobekcm_lom
xsi:schemaLocation
http:www.loc.govstandardsmetsmets.xsd
http:www.fcla.edudlsmddaitssdaitss.xsd
http:www.loc.govmodsv3mods-3-4.xsd
http:digital.uflib.ufl.edumetadatasobekcmsobekcm.xsd
METS:metsHdr CREATEDATE 2023-05-10T13:43:11Z ID LASTMODDATE 2023-05-10T10:58:43Z RECORDSTATUS COMPLETE
METS:agent ROLE CREATOR TYPE ORGANIZATION
METS:name UF,University of Florida
OTHERTYPE SOFTWARE OTHER
Go UFDC - FDA Preparation Tool
INDIVIDUAL
UFAD\renner
METS:dmdSec DMD1
METS:mdWrap MDTYPE MODS MIMETYPE textxml LABEL Metadata
METS:xmlData
mods:mods
mods:abstract lang en Food insecurity from climate change, human activity, and population growth is a global issue, affecting mostly those in developing areas. Cowpea (Vigna unguiculata [L.] Walp.) is a drought-tolerant, warm-season legume that is a source of food security in areas such as Africa, the Americas, and Asia. The purpose of this project was to develop mutant lines of dual edible-ornamental cowpeas. Sixty seeds were soaked in varying treatments of ethyl methanesulfonate (EMS) while agitated for 3 or 4 hours. Seeds were planted in sand-soil media and placed in a grow chamber with 12 hours of light, Day/Night temperatures of 27C/22C for 3 months. ANOVA models were fitted in R to compare treatments. The highest EMS concentration (80 mM) resulted in shorter plants compared to all other treatments. The water control had thicker stems than EMS treated. Lower EMS concentrations (10-20 mM) had reduced SPAD values compared to EMS 40, 80 mM, and the controls. EMS 40 mM produced the highest number of pods and SPAD values similar to controls. Seeds were harvested from each treatment level and progeny will be evaluated in future studies to monitor phenotypic changes with the goal of developing dual-purpose cowpea cultivars.
mods:accessCondition Copyright Rebecca Olivia Arias. Permission granted to the University of Florida to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
mods:language
mods:languageTerm type text English
code authority iso639-2b eng
mods:location
mods:physicalLocation University of Florida
mods:name
mods:namePart Arias, Rebecca Olivia
mods:role
mods:roleTerm creator
mods:note Awarded Bachelor of Science, magna cum laude, on April 29, 2022. Major: Plant Science. Emphasis/Concentration:
College or School: Agricultural and Life Sciences
Advisor: Esteban Fernando Rios. Advisor Department or School: Agronomy. Advisor: . Advisor Department or School:
mods:originInfo
mods:publisher University of Florida
mods:dateIssued 2023
mods:recordInfo
mods:recordIdentifier source sobekcm AA00092429_00001
mods:recordContentSource University of Florida
mods:relatedItem series
mods:part
mods:detail Year
mods:caption 2023
mods:subject
mods:topic Undergraduate Honors Thesis/Project
mods:titleInfo
mods:title Advancement of Cowpea Into a Dual Edible and Ornamental Crop
mods:typeOfResource text
DMD2
OTHERMDTYPE SOBEKCM SobekCM Custom
sobekcm:procParam
sobekcm:Aggregation ALL
UFIR
IUF
UFHONORS
UNDERGRADWORKS
sobekcm:MainThumbnail Arias_Rebecca_Honors_Thesisthm.jpg
sobekcm:Wordmark UFIR
sobekcm:Tickler 2231_SPRING_2023_HONORS
sobekcm:bibDesc
sobekcm:BibID AA00092429
sobekcm:VID 00001
sobekcm:Publisher
sobekcm:Name University of Florida
sobekcm:Source
sobekcm:statement UF University of Florida
sobekcm:SortDate 738520
METS:amdSec
METS:digiprovMD DIGIPROV1
DAITSS Archiving Information
daitss:daitss
daitss:AGREEMENT_INFO ACCOUNT PROJECT UFDC
METS:techMD TECH1
File Technical Details
sobekcm:FileInfo
METS:fileSec
METS:fileGrp USE reference
METS:file GROUPID G1 PDF1 applicationpdf CHECKSUM dfa041886b88acee05891ff30810878c CHECKSUMTYPE MD5 SIZE 1219295
METS:FLocat LOCTYPE OTHERLOCTYPE SYSTEM xlink:href Arias_Rebecca_Honors_Thesis.pdf
G2 TXT2 textplain
Arias_Rebecca_Honors_Thesis_pdf.txt
G3 METS3 unknownx-mets 8d1f923cb5f9e9154f5e68f784e28e1f 6926
AA00092429_00001.mets
METS:structMap STRUCT2 other
METS:div DMDID ADMID ORDER 0 main
ODIV1 1 Main
FILES1 Page
METS:fptr FILEID
FILES2 2
FILES3 3
PAGE 1
1 Advancement of Cowpea I nto a D ual Edible and Ornamental C rop Rebecca Arias1*; Esteban Rios, Ph.D.1 1Agronomy Department, College of Agricultural and Life Sciences, University of Florida *Corresponding Author; email: arias.rebeccaplnt@gmail.com Abstract Food insecurity from climate change, human activity, and population growth is a global issue, affecting mostly those in developing areas. Cowpea ( Vigna unguiculata [L.] Walp.) is a droughttolerant, warm season legume that is a source of food security in areas such as Africa, the Americas, and Asia. The purpose of this project was to develop mutant lines of dual edible -orna mental cowpeas. Sixty seeds were soaked in varying treatments of e thyl methanesulfonate (EMS) while agitated for 3 or 4 hours. Seeds were planted in sand-soil media and placed in a grow chamber with 12 hours of light, Day/Night temperatures of 27C/22C for 3 months. ANOVA models were fitted in R to compare treatments . The highest EMS concentration (80 mM) resulted in shorter plants compared to all other treatments. The w ater control had thicker stems than EMS treated. Lower EMS concentrations (10 -20 mM) had reduced SPAD values compared to EMS 40, 80 mM, and the controls. EMS 40 mM produced the highest number of pods and SPAD values similar to controls . Seeds were harvested from each treatment level and progeny will be evaluated in future studies to monitor phenotypic changes with the goal of developing dual-purpose cowpea cultivars. Keywords : Cowpea, Ethyl methanesulfonate, mutagenesis, plant breeding Introduction Cowpea biology and production Cowpea ( Vigna unguiculata [L.] Walp.) is a drought-tolerant legume whose origins lie in the African continent. It has a key role as a food source in SubSaharan Africa due to its ability to fix nitrogen in arid conditions, and high protein content (Moray et al., 2015). Cowpea foliage and seeds are edible both for humans and livestock. A majority of global yield comes from Africa (84%) with Nigeria being the largest worldwide producer (Kebede & Bekeko, 2020), although it is also significantly cultivated in Asia, Australia , the Caribbean, and the Americas (Pratap, 2020). Developing countries and subsistence farmers across the globe rely on pulses and legumes such as cowpea for this trifecta of food security, animal feed, and nitrogen fixation.
PAGE 2
2 The multipurpose nature of this crop makes it an attractive, versatile addition to community gardens, farms, and home growers. Increased efficiency of growing space, chemical applications, and labor costs are possible by using crops bred to be multifunctional. Like many other legumes, cowpea is a n obligate selfpollinating crop with little outcrossing . Attempts at producing crosses by hand pollination often result in the abortion of seeds, pod drops, and low seed production. These factors hinder breeders’ efforts and have slowed the overall improvement of cowpea (Boukar et al., 2020). Mutagenesis provides an alternative to traditional breeding methods, with the added benefit of assisting efforts to increase genetic diversi ty in crops that suffer from genetic bottlenecks. Agronomic significance of cowpea Cowpea is a legume native to Africa, and is a crop of specific importance to the subSaharan region – cowpeas’ primary source of diversity. It is one of the most important legume crops in the continent, where it provides nutritional security, livestock fodder, and increases in soil fertility via nitrogen fixation. It is a source of income for both farmers and food vendors (Fatokun et al., 2018), helping to keep currency flowing within their respective local economies. Cowpea is more drought tolerant than similar crops and is able to withstand lower fertility soils and overall lesser inputs. Indeed; breeders are interested in developing both drought tolerance and nitrogen fixation to breed superior lines of cowpea ( Foyer et al., 2016) . In terms of nutrition, it is high in protein, ranging from 23% to 32% of seed weight (Timko & Singh), providing the complimentary nutritional profile to many grains and tubers grown in the region as staple crops, which are typically low in protein content. For this reason, cowpea and other legumes are often called the ‘poor man’s meat’. Cowpea is often referred to as a grain, from the understanding that the seed is the most important part of the plant for human consumption. In terms of nutrition, the seed s contain vitamins and minerals such as lysine, folic acid, Vitamins A & B, thiamine, calcium, potassium, phosphorus, iron, zinc, and other trace elements (Foyer et al., 2016). In particular, folic acid (a type of B vitamin) is necessary for women during pregnancy to avoid major birth defects in the fetus’s brain and spine. According to the Centers for Disease Control and Prevention (CDC), 400 mcg (micrograms) of fol ic acid is needed daily for pregnant women ( http://www.cdc.gov/ncbddd/folicacid/ ). Folate, which is the
PAGE 3
3 naturally occurring form of folic acid according to the CDC, is found in cowpeas, where 100g of cooked cowpea provides 208 mcg of folate (Affrifah et al., 2022). Preference for cowpea preparation varies depending on the area; some prefer the beans cooked, while others prefer fresh seeds and green pods. In addition, the leaves are consumed dried or fresh in meals and provide additional nutrition, similar to the use of other greens (Tarawali et al. 1997, 2002). However, data on the fresh bean, pod, and leaf consumption are scarce as dry cowpea is typically the only data collected . Dry cowpea production globally in 2021 was estimated to be 8,986,191.25 tonnes as per the Food and Agriculture Organization of the United Nations (FAO) ( https://www.fao.org/faostat ), with the top 3 global producers being Nigeria, Niger, and Burkina Faso . Production of cowpea is expected to rise, as it is a crop able to withstand drought stress which is a more common occurrence with climate change. Mutagenesis and the use of e thyl methanesulfonate in plant breeding Traditional breeding methods have proved to be difficult, as cowpea is an obligate selfpollinating crop. Additionally, the wide origin of diversity possibly resulted in genetic drift, and therefore incompatibilities between genotypes. The flowers do not take well to the manipulation required for cross -pollination, and often times abortion of pods is the result (Boukar et al., 2020). W ith this in mind, breeders have additional tools at their disposal; mutagens. A mutation is a change in a DNA sequence; this can be deletions, insertions, substitutions – anything from a single base pair change to something as large as chromosome deletions or doubling. A spontaneous mutation is a type of mutation that occurs due to errors in DNA replication, repair, and /or recombination. In addition, the movement of genetic elements and randomly occurring DNA damage can also cause spontaneous mutations. Quantifying the number of spontaneous mutations that occur within a cell during its lifetime gives a metric called the mutation rate (Foster, 2006). In Arabidopsis thaliana, the model organism used for plants, the mutation rate under normal conditions has been estimated at 7 10 base substitutions per site per generation (Belfield et al., 2021). Spontaneous mutations are a normal part of these processes and are the source of variation that then leads to speciation by the forces of evolution. In plant breeding, these spontaneous mutations are the basis for selections that led to the development of domesticated crops. As technology progressed, scientists were able to increase that rate of mutation via mutagenesis. This was first documented by Stadler in 1928, who
PAGE 4
4 showed the possibility of inducing mutations in maize with X rays (Stadler, 1928). The methods for mutagenesis have increased since then. These range from various chemical mutagens such as hydrogen fluoride (HF) and methyl methanesulfonate (MMS), to Gamma, UV, and X ray radiation (Chaudhary et al., 2019). Plant breeders have and continue to use mutagenesis for crop improvement, increasing the genetic diversity of crops, and as a method to better understand the biological processes of plants. Ethyl methanesulfonate (EMS) is the most commonly used alkylating agent for mutagenesis in plant breeding. The majority of mutations generated by EMS are point mutations, which are almost exclusively Guanine/Cytosine to Adenine/Thymine changes (Gr eene et al., 2003) . The combination of EMS’ low cost, high availability, and mutagenic efficiency make it accessible and viable to a wide range of plant breeders ( Melsen et al., 2021) . Cowpea is a crop of interest for plant breeding, as historically it is more recently domesticated compared to crops such as corn ( Zea mays L.) , rice ( Oryza sativa L.) , and wheat ( Triticum aestivum L.) . In addition, it has characteristics that lend it to be ing a multipurpose crop; it is a legume seed grain for human consumption, animal fodder, and a method of improving soil quality and structure (Dareus et al., 2021). With the increase of home and community gardens, crops that can serve dual purposes become exceedingly attractive as individuals face space and time restrictions. Focusing on the improvement of the ornamental qualities of cowpea would elevate the crop to one that not only provides nutritional benefits but also has mental and physical benefits. T here has been increasing evidence that gardening and exposure to nature have benefits for an individual’s mental health and well being. In particular, flowering plants provided a higher benefit than foliage plants (Zhang et al., 2021). Examples of other cultivated crops with ornamental value include borage ( Borago officinalis L. ) (Tewari, Bawari, Patni, & Sah, 2019) , chards ( Beta vulgaris subsp. vulgaris ), nasturtiums ( Tropaeolum spp. L. ), peppers ( Capsicum spp.; C. annum , C. chinense , C. frutescens , and others), rhubarb ( Rheum hybridum ), and many others. Therefore, expanding breeding efforts to include more crops would give grow ers more choices in their planting configurations. Objectives The objective of this project was to develop cowpea into a multifaceted crop, focusing on edible and ornamental qualities. The intent behind this was to provide smaller stakeholders with
PAGE 5
5 a food crop that would also increase their quality of life by additional beauty and ornamental value. Materials and Methods For the purpose of this study, a selection of cowpea accessions were screene d for traits of interest in the field, with a single line (#75) being selected for the purpose of this experiment. As shown in Figures 1, 2, & 3. The combination of a bush habit, pods that curled and turned purple as they matured, and pods raised high above foliage made line 75 an attractive starting point for the development of a dual edible and ornamental crop. The biological material for this experiment were cowpea seeds of line 75, and seed were increased prior to the start of the study. Figure 2 is a photo of one such plant used for the seed increase, while Figure 3 exhibits flowers and immature pods. The chemical mutagen used was ethyl methanesulfonate (E MS ) at different concentrations to induce mutagenesis, which respectively were 10, 20, 30, 40, and 80 mM. The control was treated with DI water, and all concentrations had two treatment exposure times, 3 or 4 hours, leading to a total of 12 treatments and 5 seeds per treatment were used as replicates . The protocol for mutagenesis of cowpea seeds was adapted and modified from Opoku et al (2022) to fit the scale and scope of this experiment. Seeds were placed in a 100 mL beaker with 80 mL of DI water and agitated every 15 minutes. Seeds were then separated into 50 mL falcon tubes, with 7.5 mL of DI water and differing volumes of EMS pipetted according to treatment concentrations. These aqueous solutions corresponded to 0, 10, 20, 30, 40, and 80 mM of EMS ( Table 1 ) . Falcon tubes were placed in a rack, and then all placed in a shaker set to 200 RPM. Odd numbered treatments were left in aqueous solutions for 3 hours, even numbered treatments for 4 hours. Seeds were rinsed 3 times with DI water before being planted in a sterile 1:1 sand to soil media in black cones. The planting media had been sterilized by autoclave. The cones were set in 3 trays, in a randomized complete block design. Trays were transferred to local grow chamber facility, and then placed in a grow chamber kept at 27 C daytime and 22 C nighttime temperatures. Day length was set at 12 hours, with grow lights set to mimic daylight.
PAGE 6
6 Figure 1 . C owpea line 75 grown in a field trial at the Plant Science & Research Education Unit in Citra, Florida in Fall 2021. Line 75 was selected as it has a phenotype of interest for ornamental quality. Figure 2. Gallon sized pot of cowpea line 75, which was selected for mutagenesis due to phenotypic traits of interest.
PAGE 7
7 Figure 3. Close up of cowpea line 75’s flowers and pods. These features were of primary interest in the selection of a line for mutagenesis. Over the course of 3 months, plants were grown and observed. Height measurement in cm from soil level to the highest node was taken twice a week. Stem width was taken with an electronic caliper weekly, with values resulting from an average of 3 measurements per plant. Once true leaves were developed, a Soil Plant Analysis Development (SPAD) chlorophyll meter was used weekly. For this, 3 leaves and/or leaflets had 3 measurements each taken for a total of 9 measurements per plant that were then averaged. Reproductive stage observations were taken twice a week on bud, flower, and pod numbers. At each observation, the number of buds, flowers, and pods were documented, which showed the speed of development from bud to flower, to pod. At the end of the experiment, seed pods were counted and collected, with pods separated by branch per plant due to the possibility of chimeric growth.
PAGE 8
8 Table 1. Table of Ethyl methanesulfonate treatments of cowpea seeds, the concentration of EMS per treatment, the duration of exposure, and the volumes used to make the aqueous solution. D ata was analyzed following ANOVA in R (R Core Team, 2021) using the package agricolae ( de Mendiburu , 2021) , and graphs were created with the package ggplot2 ( Wickman, 2016). Results & Discussion Significant variation for EMS treatment was estimated via rounded calculation for number of pods, while none of the factors were signicant for bud and flower numbers (Table 2). There was no significant variation in the number of flowers across treatments a nd mean values ranged between 1.57 to 3.50. Similarly , no significant differences were detected for the number of buds and mean values ranged between 5.38 and 8.67. The number of pods increased with the EMS concentration up to a peak of 40 mM , with a decrease in the number of pods at 80 mM, and 40 mM produced significantly more pods than the water control (Figure 4). The 10 mM EMS EMS concentration in mM (mmol/L) Duration in hours Number of seeds treated per treatment Volume of solution used to treat seeds Volume of EMS in solution Control 1 0 3 5 7.5mL 0 L
PAGE 9
9 treatment had the lowest number of pods counted. These comparisons suggest that there is an ideal amount of treatment that induces higher pod production, and excess of that value reduces pod production. L ine 75 has great potential to be developed as a dual purpose crop because it produces a large number of pods, which can be eaten green or harvested for grain. Table 2. Mean square for the number of cowpea pods, number of buds, and number of flowers produced. Pod number Bud number Flower number Replicate 138.91* 13.78 ns 1.6 ns EMS Treatment 249.59** 10.45 ns 2.5 ns Time 17.35 ns 7.18 ns 0.03 ns EMS Treatment x :Time 40.25 ns 5.67 ns 8.19 ns Residuals 43 8.65 37.17 Ns: nonsignificant at P<0.05; * significant at P <0.05; ** significant at P<0.001 EMS treatment was statistically significant for height, stem width and SPAD . The difference in duration of mutagen treatment was also significant for SPAD (Table 3). Plant height was statistically similar for the water control and EMS treatments from 10 to 40 mM, and plants receiving the treatment 80 mM were the shortest (Figure 5) . Mutagenic treatments are usually deleterious in plants, and the highest EMS concentration resulted in shorter plants. Similarly, stem width was thinner in plants receiving 80 mM EMS, as well as in plants receiving 10 mM EMS. Nevertheless, despite the statist ical significance, EMS 10 and 80 MM reduced stem width only by 1 m m compared to the control (Figure 6) . Higher EMS concentrations (30 to 80 mM) had similar SPAD values to the water control, while 10 and 20 mM EMS had lower values (Figure 7). Table 3. Mean square values for cowpea plant height, stem, width, and SPAD. Height Stem width SPAD Rep licate 0.252 ns 0.023 ns 8.63 ns EMS Treatment 3.291** 0.342** 56.17 ** Time 0.401 ns 0.113 ns 83.78 ** Residuals 0.727 0.066 10.64 Ns: nonsignificant at P<0.05; * significant at P <0.05; ** significant at P<0.001
PAGE 10
10 Figure 4. G raph showing the number of cowpea pods counted semiweekly comparing a control to increasing concentrations of ethyl methanesulfonate (10 to 80 mM) . Figure 5 . G raph of average cowpea plant height across a control and increasing concentrations of ethyl methanesulfonat e (10 to 80 mM) .
PAGE 11
11 Figure 6 . G raph o f average cowpea plant stem diameter in mm across a control and increasing concentrations of ethyl methansulfonate (10 to 80 mM) . Figure 7 . G raph of cowpea SPAD value averages across the control and increasing concentrations of ethyl methanesulfonate (10 to 80 mM) .
PAGE 12
12 Conclusion Plant breeders bear the responsibility of improving society’s crops, with input from consumers, farmers, growers, and the community at large. The advancement of computing power, genomic tools, and technology available has made this process increasingly eff icient – but there always remain challenges to overcome. Understanding the context of a crop, its’ history, and its’ future is key to answering the needs of those we haven’t met yet. It is not enough to look where we are, but to look where we are going – a nd the world is getting hotter, dryer, and more crowded than ever. That is why we need crops that can handle heat, drought, but also provide multiple values while in the same space. Our arable land is not going to get bigger, so we must further maximize th e space we have. Investing in research to produce dual and multipurpose lines of crops is how we will be able to increase the value of our planting area. And with cowpea, mutagenesis has been shown to be the way to increase genetic diversity and be the backbone of developing a breeding program. T his study found that the treatment 40 mM EMS resulted in a greater number of pods and similar plant architecture to the water control, and it will be recommended to develop new cowpea breeding lines for dual-purpose applications. Using EMS treated seeds, this study observed the development of 60 plants over the course of around 3 months. Seeds were collected f rom putatively mutagenic plant s and shall be kept for future work in the UF breeding program, where the progeny of possible mutants will be grown and observed for traits of interest. Acknowledgments We would like to thank all the members and staff of the Forage Breeding and Genetics Lab at the University of Florida, Gainesville, FL for providing the seed for this experiment, assisting with data analysi s, and providing support throughout the duration of this research . We also would like to thank the College of Agricultural and Life Sciences and the Center for Undergraduate Research at the University of Florida for their patronage of this project, and assistance with all technical queries. References Affrifah, N. S., Phillips, R. D., & Saalia, F. K.. (2022). Cowpeas: Nutritional profile, processing methods and products—A review. Legume Science, 4(3). https://doi.org/10.1002/leg3.131
PAGE 13
13 Belfield, E. J., Brown, C., Ding, Z. J., Chapman, L., Luo, M., Hinde, E., . . . Harberd, N. P. (2021). Thermal stress accelerates Arabidopsis thaliana mutation rate. Genome Research , 31(1), 40-50. https://doi.org/10.1101/gr.259853.119 Boukar O, Abberton M, Oyatomi O, Togola A, Tripathi L and Fatokun C (2020) Introgression Breeding in Cowpea [Vigna unguiculata (L.) Walp.]. Front. Plant Sci. 11:567425. doi: 10.3389/fpls.2020.567425 Chaudhary, J., Deshmukh, R., & Sonah, H. (2019). Mutagenesis Approaches and Their Role in Crop Improvement. Plants , 8 (11), 467. https://doi.org/10.3390/plants8110467 de Mendiburu, F (2021). agricolae: Statistical Procedures for Agricultural Research. R package version 1.35. https://CRAN.Rproject.org/package=agricolae Fatokun, C., Girma, G., Abberton, M., Gedil, M., Unachukwu, N., Oyatomi, O., Yusuf, M., Rabbi, I., & Boukar, O.. (2018). Genetic diversity and population structure of a mini core subset from the world cowpea (Vigna unguiculata (L.) Walp.) germplasm collection. Scientific Reports, 8(1). https://doi.org/10.1038/s41598-018-34555-9 Foster, P. L. (2006). Methods for Determining Spontaneous Mutation Rates. In DNA Repair, Part B (pp. 195-213). Elsevier. https://doi.org/10.1016/s0076-6879(05)09012-9 Foy er, C. H., Lam, H.M., Nguyen, H. T., Siddique, K. H. M., Varshney, R. K., Colmer, T. D., Cowling, W., Bramley, H., Mori, T. A., Hodgson, J. M., Cooper, J. W., Miller, A. J., Kunert, K., Vorster, J., Cullis, C., Ozga, J. A., Wahlqvist, M. L., Liang, Y., Shou, H., Considine, M. J.. (2016). Neglecting legumes has compromised human health and sustainable food production. Nature Plants, 2(8), 16112. https://doi.org/10.1038/nplants.2016.112 Greene, E. A., Codomo, C. A., Taylor, N. E., Henikoff, J. G., Till, B. J., Reynolds, S. H., Enns, L. C., Burtner, C., Johnson, J. E., Odden, A. R., Comai, L., & Henikoff, S. (2003). Spectrum of chemically induced mutations from a large scale reverse-geneti c screen in Arabidopsis. Genetics, 164 (2), 731– 740. https://doi.org/10.1093/genetics/164.2.731 H. Wickham. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag New York, 2016. Kebede, E., & Bekeko, Z.. (2020). Expounding the production and importance of cowpea (Vigna unguiculata (L.) Walp.) in Ethiopia. Cogent Food & Agriculture, 6(1), 1769805. https://doi.org/10.1080/23311932.2020.1769805 Melsen, K., Van De Wouw, M., & Contreras, R. (2021). Mutation Breeding in Ornamentals. HortScience , 56(10), 1154-1165. https://doi.org/10.21273/hortsci16001-21 Moray, C., Game, E. T., & Maxted, N.. (2014). Prioritising in situ conservation of crop resources: A case study of African cowpea (Vigna unguiculata). Scientific Reports, 4(1). https://doi.org/10.1038/srep05247 Opoku Gyamfi, M., Eleblu, J. S. Y., Sarfoa, L. G., Asante, I. K., OpokuAgyemang, F., & Danquah, E. Y. (2022). Induced variations of ethyl methane sulfonate mutagenized cowpea (Vigna unguiculata L. walp) plants. Frontiers in plant science, 13, 952247. https://doi.org/10.3389/fpls.2022.952247
PAGE 14
14 Paudel, D., Dareus, R., Rosenwald, J., Muoz Amatrian, M., & Rios, E. F. (2021). Genome Wide Association Study Reveals Candidate Genes for Flowering Time in Cowpea (Vigna unguiculata [L.] Walp.). Frontiers in genetics, 12, 667038. https://doi.org/10.3389/fgene.2021.667038 Pratap, Aditya & Gupta, Sanjeev. (2020). The Beans and the Peas: From Orphan to Mainstream Crops. R Core Team (2021). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R -project.org/ Stadler, L. J. (1928). Genetic Effects of X Rays in Maize. Proceedings of the National Academy of Sciences , 14(1), 69-75. https://doi.org/10.1073/pnas.14.1.69 Tarawali SA, Singh BB, Peters M, Blade SF (1997) Cowpea haulms as fodder. In: Singh BB, Mohan Raj DR, Dashiell KE, Jackai LEN (eds) Advances in Cowpea Research. CopublicationIntl Inst Tropical Agric (IITA) and Japan Intl Res Center Agric Sci (JIRCAS). Sayce, Devon, UK, pp. 313– 325 Tarawali SA, Singh BB, Gupta SC, Tabo R, Harris F, et al. (2002) Cowpea as a key factor fo r a new approach to integrated crop –livestock systems research in the dry savannas of West Africa. In: Fatokun CA, Tarawali SA, Singh BB, Kormawa PM, M Tamo (eds) Challenges and Opportunities for Enhancing Sustainable Cowpea Production. Intl Inst Tropical Agric, Ibadan,Nigeria, pp. 233– 251 Tewari, D., Bawari, S., Patni, P., & Sah, A. N. (2019). Chapter 3.7 Borage (Borago officinalis L.). In S. M. Nabavi, & A. S. Silva (Eds.), Nonvitamin and Nonmineral Nutritional Supplements (pp. 165170). Academic Press. doi:https://doi.org/10.1016/B978-0-12-812491-8.00023-0 Timko, M. P., & Singh, B. B.. (n.d.). Cowpea, a Multifunctional Legume (pp. 227–258). https://doi.org/10.1007/978-0-387-71219-2_10 Zhang, X., Zhang, Y., & Zhai, J. (2021). Home Garden With Eco Healing Functions Benefiting Mental Health and Biodiversity During and After the COVID-19 Pandemic: A Scoping Review. Frontiers in public health, 9, 740187. https://doi.org/10.3389/fpubh.2021.740187
|
