Genetic Diversity and Population Structure Analysis in Early Generations Maize Inbreds Derived From Local Germplasm of Eastern Himalayan Regions Using Microsatellite Markers

North- Eastern parts of India fall under the Eastern Himalayan region and it is a diversity hotspot of many crops, including maize. Maize is an important traditional cereal crop grown in hill ecology of the region mainly for food, fodder and feed. To tap the potentiality of maize genetic resources in crop improvement programmes, assessment of genetic diversity is a basic requirement. Hence, in the present study, assessment of genetic diversity in thirty early generation maize inbreds developed from different germplasm of NE India was taken up using genome wide distributed fifty two microsatellite markers. The marker analysis revealed a large variation with a total of 189 alleles with an average of 3.63 alleles per marker locus. The allele size ranged from 50 bp ( phi 036 ) to 295 bp ( p 101049 ) which revealed a high level of genetic diversity among the loci. The PIC value ranged from 0.17 ( umc 1622 ) to 0.76 ( umc 1153 ) with an average value of 0.49. The value of expected Heterozygosity (H Exp ) ranged from 0.19 to 0.80 with an average of 0.57, whereas the Observed Heterozygosity (H Obs ) ranged from 0 to 0.89 with a mean of 0.14.The genetic dissimilarity between the genotype pairs ranged from 0.40 to 0.64 with a mean value of 0.57. Cluster analysis resolved the inbreds into three distinct sub-clusters. Similarly, population structure analysis also classified the inbred lines into three-subpopulations. Marker-trait associations showed a total of twelve SSR markers significantly associated with seven agronomic traits. From the present study, wide genetic variability was found among the maize inbreds with high potential to contribute new beneficial and unique alleles in genetic enhancement program of maize in India and particularly, in NE region.

Variation of Macro- and Microelements, and Trace Metals in Spring Wheat Genetic Resources in Siberia

Western Siberia is one of the major spring wheat regions of Russia, cultivating over 7 Mha. The objective of the study was to evaluate the variation of macro- and microelements, and of trace metals in four distinct groups of genetic resources: primary synthetics from CIMMYT (37 entries), primary synthetics from Japan (8), US hard red spring wheat cultivars (14), and material from the Kazakhstan–Siberian Network on Spring Wheat Improvement (KASIB) (74). The experiment was conducted at Omsk State Agrarian University, using a random complete block design with four replicates in 2017 and 2018. Concentrations of 15 elements were included in the analysis: macroelements, Ca, K, Mg, P, and S; microelements, Fe, Cu, Mn, and Zn; toxic trace elements, Cd, Co, Ni; and trace elements, Mo, Rb, and Sr. Protein content was found to be positively correlated with the concentrations of 11 of the elements in one or both years. Multiple regression was used to adjust the concentration of each element, based on significant correlations with agronomic traits and macroelements. All 15 elements were evaluated for their suitability for genetic enhancement, considering phenotypic variation, their share of the genetic component in this variation, as well as the dependence of the element concentration on other traits. Three trace elements (Sr, Mo, and Co) were identified as traits that were relatively easy to enhance through breeding. These were followed by Ca, Cd, Rb, and K. The important biofortification elements Mn and Zn were among the traits that were difficult to enhance genetically. The CIMMYT and Japanese synthetics had significantly higher concentrations of K and Sr, compared to the local check. The Japanese synthetics also had the highest concentrations of Ca, S, Cd, and Mo. The US cultivars had concentrations of Ca as high as the Japanese synthetics, and the highest concentrations of Mg and Fe. KASIB’s germplasm had near-average values for most elements. Superior germplasm, with high macro- and microelement concentrations and low trace-element concentrations, was found in all groups of material included.

Performance of Putative Mutants and Genetic Parameters of Plectranthus amboinicus (L.) through Mutation Induction With Colchicine

Genetic enhancement in vegetatively propagated crops can be done through mutation induction. Colchicine-induced mutation is one of the methods that can be employed to increase plant genetic diversity. This study aimed to determine the effect of colchicine on the performance and genetic parameters of MV3 generation of Plectranthus amboinicus (L.). This study was conducted at the Laboratory of Agricultural Seed Management, Plantation Research and Development Center, from June 2018 to June 2019. Nodes and shoots were used as explants. Mutation induction was performed using colchicine at concentrations of 0%, 0.02%, 0.04%, and 0.06%. Explant regeneration and subculture were done on MS0 medium. The number of plantlets yielded were 59 (0% concentration of colchicine), 60 (0.02%), 81 (0.04%), and 80 plantlets (0.06%), respectively. Results indicated that colchicine-induced mutation in an in vitro culture was able to generate high genetic diversity in both quantitative and qualitative characters of the plantlets. At the concentration of 0.04%, colchicine produced the highest frequency of putative mutants (28.4%). Genetic parameters in MV3 generation of P. amboinicus plantlets showed that five quantitative characters, i.e. plantlet height, number of leaves, number of shoots, leaf length, and number of roots had high heritability values at a concentration around the LC50 value (0.0275%).

CRISPER/cas9-Human Germline Gene Editing: Study of Islamic Ethical and Scientific Perspectives

The advancement of technology in medical science has just changed human lives, as well as biomedical innovations are making human lives better but lesser harmful. In past, scientifically and religiously approved techniques such as testtube baby & human stem cells therapy have served humanity especially infertile and connubial parents. Nowadays, the advancement in CRISPER/cas9 technology which is about human germline gene editing, just rekindled the religious and ethical concerns, especially in Islamic perspectives. Although, human germline genome editing and modification have been started decades ago claims about disease prevention strategies have raised many religious concerns such as tampering with God’s creation, human dignity, safety and efficacy of the technology, and human genetic enhancement. This kind of editing might result in inheritable changes in the human genome. So, questions about its status whether it should be allowed or not, need deep & serious study from religious and ethical perspectives. This study will encompass Islamic perspectives on these concerns in the light of ethical principles of Islam while considering and assessing the permissibility or lawful status of CRISPR/Cas9 mediated human germline gene editing. This research study also aims to address the controversial discussions among Muslim jurists regarding human germline gene editing as well as to comprise the related ethical regulations and concerns.

Enhancing Autonomy, Authenticity And Selecting The Child With The Best Moral Life

In the human enhancement literature, there is a recurrent fear that biomedical technologies will negatively impact the autonomy and authenticity of moral agents, even when the agents would end up having better capacities and an improved life with the aid of these technologies. I will explore several ways in which biomedical enhancement may improve the autonomy of moral agents and try to show that biomedical methods are, all things considered, beneficial to our autonomy and authenticity. I will argue that there are instances when it’s desirable to limit the autonomy of moral agents and that strict regulations are to be put in place if a great number of people will have easy access to powerful, genetic-altering technologies which can impact the life of future children. I will advocate for using assisted reproductive technologies in order to select the child with the best chance of the best moral life and in doing so I will analyse several procreative principles which have been proposed by different scholars in the genetic enhancement debate and try to determine which one would be best to adhere to. Usually, people place high value on the concept of autonomy and there are many cases in which they end up overestimating autonomy in relation to other moral values. While autonomy is important, it’s also important to know how to limit it when reasonable societal norms require it. Sometimes autonomy is defined in strong connection with the concept of authenticity, in the sense that it’s not sufficient for our choices to be autonomous if they are not also authentic. I will try to defend the idea that authenticity can be enhanced as well with the aid of enhancement technologies which can actually prove beneficial in our quest to improve our own self.

CRISPR-enhanced human adipocyte browning as cell therapy for metabolic disease

Obesity and type 2 diabetes are associated with disturbances in insulin-regulated glucose and lipid fluxes and severe comorbidities including cardiovascular disease and steatohepatitis. Whole body metabolism is regulated by lipid-storing white adipocytes as well as “brown” and “brite/beige” adipocytes that express thermogenic uncoupling protein 1 (UCP1) and secrete factors favorable to metabolic health. Implantation of brown fat into obese mice improves glucose tolerance, but translation to humans has been stymied by low abundance of primary human beige adipocytes. Here we apply methods to greatly expand human adipocyte progenitors from small samples of human subcutaneous adipose tissue and then disrupt the thermogenic suppressor gene NRIP1 by CRISPR. Ribonucleoprotein consisting of Cas9 and sgRNA delivered ex vivo are fully degraded by the human cells following high efficiency NRIP1 depletion without detectable off-target editing. Implantation of such CRISPR-enhanced human or mouse brown-like adipocytes into high fat diet fed mice decreases adiposity and liver triglycerides while enhancing glucose tolerance compared to implantation with unmodified adipocytes. These findings advance a therapeutic strategy to improve metabolic homeostasis through CRISPR-based genetic enhancement of human adipocytes without exposing the recipient to immunogenic Cas9 or delivery vectors.

Maize Zmcyp710a8 Mutant as a Tool to Decipher the Function of Stigmasterol in Plant Metabolism

Sterols are integral components of membrane lipid bilayers in eukaryotic organisms and serve as precursors to steroid hormones in vertebrates and brassinosteroids (BR) in plants. In vertebrates, cholesterol is the terminal sterol serving both indirect and direct roles in cell signaling. Plants synthesize a mixture of sterols including cholesterol, sitosterol, campesterol, and stigmasterol but the signaling role for the free forms of individual plant sterols is unclear. Since stigmasterol is the terminal sterol in the sitosterol branch and produced from a single enzymatic step, modifying stigmasterol concentration may shed light on its role in plant metabolism. Although Arabidopsis has been the model of choice to study sterol function, the functional redundancy of AtCYP710A genes and the presence of brassicasterol may hinder our ability to test the biological function of stigmasterol. We report here the identification and characterization of ZmCYP710A8, the sole maize C-22 sterol desaturase involved in stigmasterol biosynthesis and the identification of a stigmasterol-free Zmcyp710a8 mutant. ZmCYP710A8 mRNA expression pattern correlated with transcripts for several sterol biosynthesis genes and loss of stigmasterol impacted sterol composition. Exogenous stigmasterol also had a stimulatory effect on mRNA for ZmHMGR and ZmSMT2. This demonstrates the potential of Zmcyp710a8 in understanding the role of stigmasterol in modulating sterol biosynthesis and global cellular metabolism. Several amino acids accumulate in the Zmcyp710a8 mutant, offering opportunity for genetic enhancement of nutritional quality of maize. Other cellular metabolites in roots and shoots of maize and Arabidopsis were also impacted by genetic modification of stigmasterol content. Yet lack of obvious developmental defects in Zmcyp710a8 suggest that stigmasterol might not be essential for plant growth under normal conditions. Nonetheless, the Zmcyp710a8 mutant reported here is of great utility to advance our understanding of the additional roles of stigmasterol in plant metabolism. A number of biological and agronomic questions can be interrogated using this tool such as gene expression studies, spatio-temporal localization of sterols, cellular metabolism, pathway regulation, physiological studies, and crop improvement.

Current Perspectives on Reducing the β-ODAP Content and Improving Potential Agronomic Traits in Grass Pea (Lathyrus sativus L.)

Grass pea is well-established as one of the most resilient and versatile crops that can thrive under extreme climatic circumstances such as cold, heat, drought, salt-affected soils, submergence, and excessive rainfall along with resistance to several diseases and pests. However, despite the awareness of its virtues, its cultivation globally has decreased recently owing to the presence of a neurotoxin, β-N-oxalyl-L-α, β-diaminopropionic acid (β-ODAP), in the seedlings and seeds of this legume, which has been reported to cause neurolathyrism, a non-reversible neurological disorder in humans and animals. Significant repositories of Lathyrus germplasm are available across countries that have provided access to a wide range of agro-morphological traits as well as the low β ODAP content. Efforts have been made worldwide to use these germplasms for the genetic enhancement of grass pea to make this food safe for human consumption. Efforts on molecular breeding of this crop are also lagging. However, during the last decade, the research scenario has changed with some efforts being made toward improving this climate resilient pulse in terms of genomic resources. Molecular markers have also been used to evaluate the interspecific diversity as well as the phylogenetic relationship among the species and mapping studies. Intron-targeted amplified polymorphic, genomic simple sequence repeat, resistance genes analogs, and disease resistance markers developed for other legume species have been successfully cross-amplified in grass pea. Transcriptomic studies have recently been undertaken on grass pea by deploying several second-generation sequencing techniques. In addition, a few studies have attempted to unveil the genes and the underlying mechanism conferring biotic and abiotic stress or regulating the pathway of β-ODAP in grass pea. Proteomics has accelerated the identification studies on differential proteomes in response to salinity and low-temperature stress conditions for unveiling the common signaling pathways involved in mitigating these abiotic stresses and in discovering differentially regulated proteins. In grass pea, a metabolomics approach has been used to identify the metabolic processes associated with β-ODAP synthesis. Genome sequencing of grass pea is under way which is expected to be vital for whole-genome re-sequencing and gene annotation toward the identification of genes with novel functions. Recently, a draft genome sequence of grass pea was developed, and some efforts are underway to re-sequence a diverse panel of grass pea comprising 384 germplasm lines. Owing to the scantiness of a successful transformation protocol, research on the application of modern approaches of genome editing like the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) or CRISPR-associated protein 9 (CRISPR/Cas9) system for the engineering of signaling pathways or regulatory mechanisms seeks immediate attention to reduce the β-ODAP content in seeds and to improve the potential agronomic traits in grass pea.

Phenotyping and Association Analysis of Grain Zinc and Iron Content with Seed Yield in Diverse Local Germplasm of Rice

Background: Rice (Oryza sativa L.) feeds more than half of the world population, but it is a poor source of zinc (Zn) and iron (Fe). Therefore, there is a need for Zn and Fe- biofortified rice in the food chain. For this, rice breeding needs to be re-oriented to improve the status of grain Zn and Fe content while increasing the yield potential. Identification of micronutrient-rich donors and their association study with agro-economic traits can pave the way for nutritional and food security. Methods: Zinc and iron content of a set of 92 rice genotypes was estimated in the aliquot of seed extract by using an Inductive Coupled Plasma-Optical Emission Spectrophotometer (ICP-OES) to explore donors and to study the degree of association of these micronutrients with agro-economic traits including grain quality traits and seed yield. Result: The top Fe (≥50 ppm) and Zn dense (≥40 ppm) genotypes identified were P44 mutant selection-1, ORCZ 75-3-1, Basudha, Malliphulajhuli, Tikimahsuri and Nikipankhia. P44 mutant Sel.-1 and ORCZ 75-3-1 had good yield potential (44 q ha-1). Grains/panicles and the number of effective bearing tillers/hill maintained an appreciably strong positive association with seed yield, while plant height had an inverse relationship. Grain Fe content positively correlated with panicle length. On the other hand, Zn is positively associated with tillering ability and grains/panicle. Grain Fe and Zn revealed strong a inter se positive association. Interestingly, grain Fe revealed no association, but grain Zn revealed significant positive relationship with seed yield. This envisaged better scope for genetic enhancement of grain Zn content along with substantial increase in grain Fe without any yield penalty.