Application of SCoT markers for accessing of genetic diversity of gramineous forage grass species

Using the SCoT marker system, 8 varieties of cereal grasses belonging to 5 species were analyzed: Festuca pratensis, Lolium perenne, Lolium multiflorum, Festuca rubra, Festulolium. Of the 10 tested SCoT markers, 7 informative markers were selected that reveal interspecies genetic polymorphism. According to the results of the analysis, DNA profiles characteristic of each studied species were obtained, and primers allowing to detect intervarietal differences for subsequent identification and molecular genetic passportization were selected.

Analysis of genetic variability in F2 interspecific hybrids of mung bean (Vigna radiata) using inter-retrotransposon amplified polymorphism marker system

Fatmawati Y, Setiawan AB, Purwantoro A, Respatie DW, Teo CH. 2021. Analysis of genetic variability in F2 interspecific hybrids of mung bean (Vigna radiata) using inter-retrotransposon amplified polymorphism marker system. Biodiversitas 22: 4880-4889. Mung bean (Vigna radiata L. Wilczek) categorized as one of the pivotal annual crops of Vigna genera is commonly cultivated in rotation with the cereal crops during the drought season. Conversely, to ameliorate its stunted productivity, the interspecific hybridization technique has been introduced between the mung bean and the common bean to promote genetic improvement with the breeding projects in Indonesia. However, since mung bean is a self-pollinated crop and has a narrow genetic base, the selection and improvement of a specific trait using marker-assisted selection is more challenging. Hence, a precautionary investigation is imperative to evaluate the progenies resulting from interspecific hybridization using an ideal marker. This study aimed to investigate the genetic variability of the F2 population of the interspecific mung bean hybrids using retrotransposon-based markers, particularly Inter-Retrotransposon Amplified Polymorphism (IRAP) markers. In this study, we identified retrotransposon from the mung bean genome and determined the Long Terminal Repeat (LTR) sequence using the LTR Finder. The IRAP primers were designed from a conserved region of the LTR sequence. One hundred of the F2 interspecific hybrids generated from the crossing between mung bean and common bean were successfully discriminated by IRAP markers. The IRAP marker showed high heterozygosity and moderate Polymorphic Information Content (PIC) values. The IRAP markers were able to detect genetic variability in the F2 progenies resulting from the interspecific hybridization. Cluster analysis showed that 100 of the F2 progenies were grouped into three clusters. This study demonstrated that retrotransposon-based markers can offer an effective approach for evaluating the segregation in the F2 population of intercross hybrids in the mung bean.

ptxD/Phi as alternative selectable marker system for genetic transformation for bio-safety concerns: a review

Antibiotic and herbicide resistance genes are the most common marker genes for plant transformation to improve crop yield and food quality. However, there is public concern about the use of resistance marker genes in food crops due to the risk of potential gene flow from transgenic plants to compatible weedy relatives, leading to the possible development of “superweeds” and antibiotic resistance. Several selectable marker genes such as aph, nptII, aaC3, aadA, pat, bar, epsp and gat, which have been synthesized to generate transgenic plants by genetic transformation, have shown some limitations. These marker genes, which confer antibiotic or herbicide resistance and are introduced into crops along with economically valuable genes, have three main problems: selective agents have negative effects on plant cell proliferation and differentiation, uncertainty about the environmental effects of many selectable marker genes, and difficulty in performing recurrent transformations with the same selectable marker to pyramid desired genes. Recently, a simple, novel, and affordable method was presented for plant cells to convert non-metabolizable phosphite (Phi) to an important phosphate (Pi) for developing cells by gene expression encoding a phosphite oxidoreductase (PTXD) enzyme. The ptxD gene, in combination with a selection medium containing Phi as the sole phosphorus (P) source, can serve as an effective and efficient system for selecting transformed cells. The selection system adds nutrients to transgenic plants without potential risks to the environment. The ptxD/Phi system has been shown to be a promising transgenic selection system with several advantages in cost and safety compared to other antibiotic-based selection systems. In this review, we have summarized the development of selection markers for genetic transformation and the potential use of the ptxD/Phi scheme as an alternative selection marker system to minimize the future use of antibiotic and herbicide marker genes.

Developing a behavioural marker system for farmer non-technical skills (FLINTS).

Objective: Non-technical skills, the social and cognitive skills thought necessary for safe and effective working, have been studied within the farming context over the past six years. However, these skills are not yet taught as part of a safety curriculum for farmers, due, in part, to a lack of defined framework and assessment system. The current paper describes the development of the FLINTS behavioural marker system for discussion, observation, evaluation and feedback on non-technical skills for farmers.Method: The development of the behavioural marker system proceeded through three key stages. First, the current research knowledge on non-technical skills was synthesised to compile a list of non-technical skill categories and elements. Second, a series of discussion groups with subject matter experts was conducted to develop behavioural markers for each element. Lastly, refinement and review of the system was undertaken by academics and experts.Results: The FLINTS taxonomy containing five non-technical skill categories and 16 elements was produced. The non-technical skill categories comprised situation awareness, teamwork & communication, leadership, task management and decision-making each with specific elements and behavioural markers.Conclusion: FLINTS represents the first behavioural marker system for farmer non-technical skills, constructed through expert knowledge and advice via discussion and review groups, combined with underpinning research findings. This represents the first step towards the development of non-technical training and assessment for farmers. The FLINTS system was produced as a handbook and is freely available to all potential users (https://research.abdn.ac.uk/nts-farming/flints/).

Pose Estimation of Excavator Manipulator Based on Monocular Vision Marker System

Excavation is one of the broadest activities in the construction industry, often affected by safety and productivity. To address these problems, it is necessary for construction sites to automatically monitor the poses of excavator manipulators in real time. Based on computer vision (CV) technology, an approach, through a monocular camera and marker, was proposed to estimate the pose parameters (including orientation and position) of the excavator manipulator. To simulate the pose estimation process, a measurement system was established with a common camera and marker. Through comprehensive experiments and error analysis, this approach showed that the maximum detectable depth of the system is greater than 11 m, the orientation error is less than 8.5°, and the position error is less than 22 mm. A prototype of the system that proved the feasibility of the proposed method was tested. Furthermore, this study provides an alternative CV technology for monitoring construction machines.

Quick Identification of Genetic Provenances and Best Practice Woodland Management of Chinese Pine (Pinus tabuliformis)

Chinese pine (Pinus tabuliformis) is one of the most widespread, ecologically and economically important tree species in North China. In this study, we analyzed and compared the genetic diversity and population structure of 158 individuals from 17 populations of P. tabuliformis, by group III, a new mitochondrial marker system (nad1-2, nad4-3, nad5-1 and nad7-1) with two other groups of marker systems. In contrast to the conservation in the evolution of the mitochondrial sequence, the new mitochondrial marker system of P. tabuliformis shows as extremely high polymorphism as 25, whose haplotypes are more than four times of the group I marker system (nad1-2, nad4-3, and nad5-1) as 8 haplotypes. Although the group II, nad7-1 (19 haplotypes), showed high resolution in the provenance identification of P. tabuliformis, the new mitochondrial marker system is more accurate for detection of specific groups like HL, WT and NS and powerful to differentiate populations between GD and SS. The results suggested that the new mitochondrial marker system is as high as the resolution of GBS (genotype by seqencing). It is much more available and will be of great help to provenance identification and molecular assisted breeding of P. tabuliformis. This study will make theoretical foundation for the following studies on the evaluation, cultivation and germplasm management of P. tabuliformis populations and aid the breeding, biodiversity and conservation programs of forest species.

Magnetically Guided Localization Using a Guiding-Marker System® and a Handheld Magnetic Probe for Nonpalpable Breast Lesions: A Multicenter Feasibility Study in Japan

Accurate pre-operative localization of nonpalpable lesions plays a pivotal role in guiding breast-conserving surgery (BCS). In this multicenter feasibility study, nonpalpable breast lesions were localized using a handheld magnetic probe (TAKUMI) and a magnetic marker (Guiding-Marker System®). The magnetic marker was preoperatively placed within the target lesion under ultrasound or stereo-guidance. Additionally, a dye was injected subcutaneously to indicate the extent of the tumor excision. Surgeons checked for the marker within the lesion using a magnetic probe. The magnetic probe could detect the guiding marker and accurately localize the target lesion intraoperatively. All patients with breast cancer underwent wide excision with a safety margin of ≥5 mm. The presence of the guiding-marker within the resected specimen was the primary outcome and the pathological margin status and re-excision rate were the secondary outcomes. Eighty-seven patients with nonpalpable lesions who underwent BCS, from January to March of 2019 and from January to July of 2020, were recruited. The magnetic marker was detected in all resected specimens. The surgical margin was positive only in 5/82 (6.1%) patients; these patients underwent re-excision. This feasibility study demonstrated that the magnetic guiding localization system is useful for the detection and excision of nonpalpable breast lesions.