Growth, seed development and genetic analysis in wild type and Def mutant of Pisum sativum L

<p>Abstract</p> <p>Background</p> <p>The <it>def </it>mutant pea (<it>Pisum sativum </it>L) showed non-abscission of seeds from the funicule. Here we present data on seed development and growth pattern and their relationship in predicting this particular trait in wild type and mutant lines as well as the inheritance pattern of the <it>def </it>allele in F₂and F₃populations.</p> <p>Findings</p> <p>Pod length and seed fresh weight increase with fruit maturity and this may affect the abscission event in pea seeds. However, the seed position in either the distal and proximal ends of the pod did not show any difference. The growth factors of seed fresh weight (FW), width of funicles (WFN), seed width (SW) and seed height (SH) were highly correlated and their relationships were determined in both wild type and <it>def </it>mutant peas. The coefficient of determination <it>R</it>²values for the relationship between WFN and FW, SW and SH and their various interactions were higher for the <it>def </it>dwarf type. Stepwise multiple regression analysis showed that variation of WFN was associated with SH and SW. Pearson's chi square analysis revealed that the inheritance and segregation of the <it>Def </it>locus in 3:1 ratio was significant in two F₂populations. Structural analysis of the F3 population was used to confirm the inheritance status of the <it>Def </it>locus in F₂heterozygote plants.</p> <p>Conclusions</p> <p>This study investigated the inheritance of the presence or absence of the <it>Def </it>allele, controlling the presence of an abscission zone (AZ) or an abscission-less zone (ALZ) forming in wild type and mutant lines respectively. The single major gene (<it>Def</it>) controlling this phenotype was monogenic and <it>def </it>mutants were characterized and controlled by the homozygous recessive <it>def </it>allele that showed no palisade layers in the hilum region of the seed coat.</p

Application of Genetic Engineering for Control of Bacterial Wilt Disease of Enset, Ethiopia’s Sustainability Crop

Enset (Ensete ventricosum (Welw.) Cheesman) is one of the Ethiopia’s indigenous sustainability crops supporting the livelihoods of about 20 million people, mainly in the densely populated South and Southwestern parts of the country. Enset serves as a food security crop for humans, animal feed, and source of fiber for the producers. The production of enset has been constrained by plant pests, diseases, and abiotic factors. Among these constraints, bacterial wilt disease has been the most important limiting factor for enset production since its outbreak five decades ago. There is no known bacterial wilt disease resistant genetic material in the enset genetic pool to transfer this trait to susceptible enset varieties through conventional breeding. Moreover, the absence of effective chemicals against the disease has left farmers without means to combat bacterial wilt for decades. Genetic engineering has been the alternative approach to develop disease resistant plant materials in other crops where traditional breeding tools are ineffective. This review discusses enset cultivation and recent developments addressing the control of bacterial wilt disease in enset and related crops like banana to help design effective strategies

Next generation sequencing as a method to verify virus elimination using heat treatment and meristem tip culture in the five most widely used sweet potato varieties in Ethiopia

Sweet potato (Ipomoea batatas L. Lam) has become one of the staple crops in Africa in the last 20 years. In Ethiopia, sweet potato is the second most widely grown root crop and is the first regarding the production per hectare. Thus, there is a great demand of planting material throughout the country. Currently, planting material is usually obtained from own previous season harvest, local markets or from the neighboring fields since no certified clean planting material production scheme has been established in Ethiopia yet. Unfortunately, this practice has contributed to the spread of viral diseases throughout the country. Elimination of viruses from infected plants is a tedious job, which requires efficient methods to eliminate the virus and also to verify that the plants are indeed virus-free. In the case of sweet potato, it was observed that heat treatment, combined with meristem tip culture is an efficient method for virus elimination. Previous findings indicate that reverse transcription (RT) PCR is more efficient than ELISA to verify the efficiency of virus elimination. In this study, the use of next generation sequencing (NGS) was explored as a verification method and compared with RT-PCR. The results show that NGS seems to be more efficient than RT-PCR, although also prone to inconclusive results.publishedVersio

An update of sweet potato viral disease incidence and spread in Ethiopia

Sweet potato (Ipomoea batatas (L.) Lam.) is an important root crop for poor farmers in developing countries. Since the late 1980s, viral diseases have increasingly become a threat to sweet potato production in Ethiopia. This review paper presents the role of sweet potato production for ensuring food security, the level of sweet potato virus research, including the types of viral species identified and their current level of incidences in Ethiopia. Sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV), Sweet potato virus 2 (SPV2), Sweet potato virus G (SPVG), and Cucumber mosaic virus (CMV) were reported in Ethiopia, where the first two are the most common and exist at high incidences. In addition, this paper discusses the virus vectors, virus transmission methods to new farms, factors exacerbating the rate of viral incidence and the methods used to reduce the incidences. Moreover, it highlights methods of sweet potato viruses’ detection and cleaning of infected materials in use and the challenges encountered towards the efficient utilization of the methods. Finally, we suggest major intervention techniques that will integrate all key players in managing the impact of the virus on sweet potato production to improve productivity and ensuring food security in Ethiopia. The findings obtained from this review could be an input for the current research on sweet potato improvement (both planting materials and routines) in Ethiopia.publishedVersio

Immunolocalization of pectic polysaccharides during abscission in pea seeds (Pisum sativum L.) and in abscission less def pea mutant seeds

BACKGROUND: In pea seeds (Pisum sativum L.), the presence of the Def locus determines abscission event between its funicle and the seed coat. Cell wall remodeling is a necessary condition for abscission of pea seed. The changes in cell wall components in wild type (WT) pea seed with Def loci showing seed abscission and in abscission less def mutant peas were studied to identify the factors determining abscission and non-abscission event. METHODS: Changes in pectic polysaccharides components were investigated in WT and def mutant pea seeds using immunolabeling techniques. Pectic monoclonal antibodies (1 → 4)-β-d-galactan (LM5), (1 → 5)-α-l-arabinan(LM6), partially de-methyl esterified homogalacturonan (HG) (JIM5) and methyl esterified HG (JIM7) were used for this study. RESULTS: Prior to abscission zone (AZ) development, galactan and arabinan reduced in the predestined AZ of the pea seed and disappeared during the abscission process. The AZ cells had partially de-methyl esterified HG while other areas had highly methyl esterified HG. A strong JIM5 labeling in the def mutant may be related to cell wall rigidity in the mature def mutants. In addition, the appearance of pectic epitopes in two F(3) populations resulting from cross between WT and def mutant parents was studied. As a result, we identified that homozygous dominant lines (Def/Def) showing abscission and homozygous recessive lines (def/def) showing non-abscission had similar immunolabeling pattern to their parents. However, the heterogeneous lines (Def/def) showed various immunolabeling pattern and the segregation pattern of the Def locus. CONCLUSIONS: Through the study of the complexity and variability of pectins in plant cell walls as well as understanding the segregation patterns of the Def locus using immunolabeling techniques, we conclude that cell wall remodeling occurs in the abscission process and de-methyl esterification may play a role in the non-abscission event in def mutant. Overall, this study contributes new insights into understanding the structural and architectural organization of the cell walls during abscission

Next generation sequencing as a method to verify virus elimination using heat treatment and meristem tip culture in the five most widely used sweet potato varieties in Ethiopia

Sweet potato (Ipomoea batatas L. Lam) has become one of the staple crops in Africa in the last 20 years. In Ethiopia, sweet potato is the second most widely grown root crop and is the first regarding the production per hectare. Thus, there is a great demand of planting material throughout the country. Currently, planting material is usually obtained from own previous season harvest, local markets or from the neighboring fields since no certified clean planting material production scheme has been established in Ethiopia yet. Unfortunately, this practice has contributed to the spread of viral diseases throughout the country. Elimination of viruses from infected plants is a tedious job, which requires efficient methods to eliminate the virus and also to verify that the plants are indeed virus-free. In the case of sweet potato, it was observed that heat treatment, combined with meristem tip culture is an efficient method for virus elimination. Previous findings indicate that reverse transcription (RT) PCR is more efficient than ELISA to verify the efficiency of virus elimination. In this study, the use of next generation sequencing (NGS) was explored as a verification method and compared with RT-PCR. The results show that NGS seems to be more efficient than RT-PCR, although also prone to inconclusive results

Next generation sequencing as a method to verify virus elimination using heat treatment and meristem tip culture in the five most widely used sweet potato varieties in Ethiopia

Sweet potato (Ipomoea batatas L. Lam) has become one of the staple crops in Africa in the last 20 years. In Ethiopia, sweet potato is the second most widely grown root crop and is the first regarding the production per hectare. Thus, there is a great demand of planting material throughout the country. Currently, planting material is usually obtained from own previous season harvest, local markets or from the neighboring fields since no certified clean planting material production scheme has been established in Ethiopia yet. Unfortunately, this practice has contributed to the spread of viral diseases throughout the country. Elimination of viruses from infected plants is a tedious job, which requires efficient methods to eliminate the virus and also to verify that the plants are indeed virus-free. In the case of sweet potato, it was observed that heat treatment, combined with meristem tip culture is an efficient method for virus elimination. Previous findings indicate that reverse transcription (RT) PCR is more efficient than ELISA to verify the efficiency of virus elimination. In this study, the use of next generation sequencing (NGS) was explored as a verification method and compared with RT-PCR. The results show that NGS seems to be more efficient than RT-PCR, although also prone to inconclusive results

An update of sweet potato viral disease incidence and spread in Ethiopia

Sweet potato (Ipomoea batatas (L.) Lam.) is an important root crop for poor farmers in developing countries. Since the late 1980s, viral diseases have increasingly become a threat to sweet potato production in Ethiopia. This review paper presents the role of sweet potato production for ensuring food security, the level of sweet potato virus research, including the types of viral species identified and their current level of incidences in Ethiopia. Sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV), Sweet potato virus 2 (SPV2), Sweet potato virus G (SPVG), and Cucumber mosaic virus (CMV) were reported in Ethiopia, where the first two are the most common and exist at high incidences. In addition, this paper discusses the virus vectors, virus transmission methods to new farms, factors exacerbating the rate of viral incidence and the methods used to reduce the incidences. Moreover, it highlights methods of sweet potato viruses’ detection and cleaning of infected materials in use and the challenges encountered towards the efficient utilization of the methods. Finally, we suggest major intervention techniques that will integrate all key players in managing the impact of the virus on sweet potato production to improve productivity and ensuring food security in Ethiopia. The findings obtained from this review could be an input for the current research on sweet potato improvement (both planting materials and routines) in Ethiopia