Sclerotinia stem rot (Sclerotinia sclerotiorum) is a major disease of Brassica oilseeds. As suitable donors to develop resistant cultivars are not available in crop Brassicas, we introgressed resistance from a wild Brassicaceae species, B. fruticulosa. We produced 206 B. juncea-B. fruticulosa introgression lines (ILs). These were assessed for pollen grain fertility, genome size variations and resistance responses to Sclerotinia following stem inoculations under disease-conducive conditions. Of these, 115 ILs showing normal fertility and genome size were selected for cytogenetic characterization using florescent genomic in situ hybridization (Fl-GISH). B. fruticulosa segment substitutions were indicated in 28 ILs. These were predominantly terminal and located on B-genome chromosomes. A final set of 93 highly fertile and euploid (2n = 36) ILs were repeat-evaluated for their resistance responses during 2014–15. They were also genotyped with 202 transferable and 60 candidate gene SSRs. Association mapping allowed detection of ten significant marker trait associations (MTAs) after Bonferroni correction. These were: CNU-m157-2, RA2G05, CNU-m353-3, CNU-m442-5, ACMP00454-2, ACMP00454-3, EIN2-3-1, M641-1, Na10D09-1 and Na10D11-1. This is the first time such a molecular mapping technique has been deployed with introgression lines carrying genomic segments from B. fruticulosa, and the first to show that they possess high levels of resistance against S. sclerotiorum.

Plantago ovata is an economically and medicinally important plant of the family Plantaginaceae. It is used extensively for the production of seed husk for its application in pharmaceutical, food and cosmetic industries. In the present study, the transcriptome of P. ovata ovary was sequenced using Illumina Genome Analyzer platform to characterize the mucilage biosynthesis pathway in the plant. De novo assembly was carried out using Oases followed by velvet. A total of 46,955 non-redundant transcripts (≥100 bp) using ~29 million high-quality paired end reads were generated. Functional categorization of these transcripts revealed the presence of several genes involved in various biological processes like metabolic pathways, mucilage biosynthesis, biosynthesis of secondary metabolites and antioxidants. In addition, simple sequence-repeat motifs, non-coding RNAs and transcription factors were also identified. Expression profiling of some genes involved in mucilage biosynthetic pathway was performed in different tissues of P. ovata using Real time PCR analysis. The study has resulted in a valuable resource for further studies on gene expression, genomics and functional genomics in P. ovata.

We report first-time synthesis of a stable Brassica allohexaploid. It may evolve into a new species and also advance our understanding of pairing regulation and genome evolution in complex allopolyploids. Crop Brassicas include both monogenomic and digenomic species. A trigenomic Brassica (AABBCC) is not known to exist in nature. Past attempts to synthesize a stable allohexaploid were not successful due to aberrant meiosis and very high proportion of aneuploid plants in the selfed progenies. We report the development of a stable allohexaploid Brassica (2n = 54; AABBCC). Genomic in situ hybridization confirmed the complete assemblage of three genomes. Only allohexaploids involving B. rapa cv. R01 (2n = 20; AA) as pollinator with a set of B. carinata (2n = 34; BBCC) were stable. These exhibited a high proportion (0.78-0.94) of pollen mother cells with normal meiosis and an excellent hexaploid ratio (0.80-0.94) in the selfed progenies. Stability of two allohexaploid combinations was demonstrated from H to H generations at two very diverse locations in India. Graphical genotyping of allohexaploids allowed detection of chromosome fragment exchanges among three genomes. These were much smaller for meiotically stable allohexaploids as compared to unstable ones. The putative hexaploids were morphologically closer to the female donor, B. carinata, for leaf morphology, inflorescence structure and flower shape. The newly formed allohexaploid may also provide unique opportunities to investigate the immediate genetic and genomic consequences of a Brassica allohexaploid with three resident genomes.

Two intergeneric hybrids involving wild species Erucastrum cardaminoides (2 n=18, E(cd) E(cd)) and two crop brassica species, Brassica rapa (2 n=20, AA) and B. nigra (2 n=16, BB), were synthesized through in vitro sequential ovary culture. Morphological, molecular and cytological studies were conducted to establish their hybridity. Both hybrids, though morphologically distinct, were intermediate phenotypically between their respective parents. Cytological analysis of the E. cardaminoides x B. rapa hybrid (2 n=19), revealed the occurrence of 17 I+1 II at diakinesis/metaphase in the majority (28%) of the pollen mother cells (PMCs), whereas in E. cardaminoides x B. nigra hybrid (2 n=17), 13 I+2 II was the predominant (32%) meiotic configuration. A maximum of 5 II was recorded in both hybrids, indicating homoeologous pairing in the respective combined genomes. Chromosome doubling by colchicine application gave rise to two new amphiploids (AA E(cd)E(cd) and BB E(cd)E(cd)) having normal chromosome pairing and pollen fertility. The occasional occurrence of one quadrivalent in the amphiploids confirmed partial homoeology between the E(c) and A/B genomes. The E. cardaminoides x B. nigra hybrid and amphiploid appeared to be tolerant to alternaria blight under field conditions.

This paper presents the algorithms and system architecture of an autonomous racecar. The introduced vehicle is powered by a software stack designed for robustness, reliability, and extensibility. In order to autonomously race around a previously unknown track, the proposed solution combines state of the art techniques from different fields of robotics. Specifically, perception, estimation, and control are incorporated into one high-performance autonomous racecar. This complex robotic system, developed by AMZ Driverless and ETH Zurich, finished 1st overall at each competition we attended: Formula Student Germany 2017, Formula Student Italy 2018 and Formula Student Germany 2018. We discuss the findings and learnings from these competitions and present an experimental evaluation of each module of our solution.

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is a major disease of crop brassicas, with inadequate variation for resistance in primary gene pools. We utilized a wild Brassicaceae species with excellent resistance against stem rot to develop a set of B. juncea - B. fruticulosa introgression lines (ILs). These were assessed for resistance using a highly reproducible stem inoculation technique against a virulent pathogen isolate. Over 40% of ILs showed higher levels of resistance. IL-43, IL-175, IL-215, IL-223 and IL-277 were most resistant ILs over three crop seasons. Sequence reads (21x) from the three most diverse ILs were then used to create B. juncea pseudomolecules, by replacing SNPs of reference B. juncea with those of re-sequenced ILs. Genotyping by sequencing (GBS) was also carried out for 88 ILs. Resultant sequence tags were then mapped on to the B. juncea pseudomolecules, and SNP genotypes prepared for each IL. Genome wide association studies helped to map resistance responses to stem rot. A total of 13 significant loci were identified on seven B. juncea chromosomes (A01, A03, A04, A05, A08, A09 and B05). Annotation of the genomic region around identified SNPs allowed identification of 20 candidate genes belonging to major disease resistance protein families, including TIR-NBS-LRR class, Chitinase, Malectin/receptor-like protein kinase, defensin-like (DEFL), desulfoglucosinolate sulfotransferase protein and lipoxygenase. A majority of the significant SNPs could be validated using whole genome sequences (21x) from five advanced generation lines being bred for Sclerotinia resistance as compared to three susceptible B. juncea germplasm lines. Our findings not only provide critical new understanding of the defensive pathway of B. fruticulosa resistance, but will also enable development of marker candidates for assisted transfer of introgressed resistant loci in to agronomically superior cultivars of crop Brassica.

The seed husk of Plantago ovata known as psyllium (Isabgol) yields medicinally important mucilage. The amount of mucilage produced is about 25 % (by weight) of the total seed yield. In the present study, an attempt was made to increase the amount of mucilage through callus cultures of P. ovata. The first step involved establishment of callus cultures in P. ovata. Leaf explants from 10 to 20 day old seedlings were cultured on MS basal medium supplemented with various concentrations of different plant growth regulators. The highest rate of callus induction (89 %) was obtained on MS medium containing 0.5 mg l -1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg l -1 Kinetin. The mucilage content was estimated from the callus obtained in different media. The best mucilage production was obtained in the MS medium supplemented with 0.25 mg l -1 2,4-D and 0.25 mg l -1 thidiazuron. Significant differences with regard to the total mucilage content were recorded. Overall, the callus produced nearly five times more mucilage than the seeds. The present technology provides an alternative route to production of large quantities of mucilage without plants.The seed husk of Plantago ovata Forsk, is an effective laxative. Seeds of P. ovata release mucilage in water, which is an anionic polysaccharide of L-arabinose, D-xylose and Dgalacturonic acid [1], and is used to treat diverticulitis [2], constipation, diarrhoea, haemorrhoids, bladder problems, irritable bowel syndrome, high blood pressure, high cholesterol and type 2 diabetes [3]. Mucilage of plant origin is in high demand nowadays, and it is very difficult to fulfil the entire demand by field grown plants only. For this purpose, a striking and very promising alternative system for commercial exploitation is the plant tissue culture by using cell suspension culture systems [4]. However, so far any attempts to increase mucilage have not yielded positive results in P. ovata. Studies along these lines have been conducted in Plantago lanceolata where in media having different combinations of plant growth regulators (PGRs) have been used to obtain mucilage from callus [5]. It has been reported that callus cultures are relatively rich in mucilage which commonly make up between 8-10 and 0.2-3.7 % of dry weight in some plants [6,7]. To the best of our knowledge, there are no reports on in vitro mucilage production from the callus cultures of P. ovata. Therefore, the present study was aimed to explore the possibility of mucilage production through in vitro culture.Seeds of P. ovata were available in the collection center of School of Biotechnology, University of Jammu, Jammu, India. The seeds were sterilized first with 70 % V/V ethanol for 1 min, then with 5 % w/v sodium-hypochlorite solution for 20 min, finally washed five times with sterile distilled water. The sterilized seeds were aseptically germinated on an agar-solidified MS basal medium [8] and incubated at a temperature of 25 ± 1°C. Leaf segments (1.5 cm) were excised from 10 to 20 day-old seedlings and were used as explants ...

Allohexaploid Brassica populations reveal ongoing segregation for fertility, while genotype influences fertility and meiotic stability. Creation of a new Brassica allohexaploid species is of interest for the development of a crop type with increased heterosis and adaptability. At present, no naturally occurring, meiotically stable Brassica allohexaploid exists, with little data available on chromosome behaviour and meiotic control in allohexaploid germplasm. In this study, 100 plants from the cross B. carinata × B. rapa (A2 allohexaploid population) and 69 plants from the cross (B. napus × B. carinata) × B. juncea (H2 allohexaploid population) were assessed for fertility and meiotic behaviour. Estimated pollen viability, self-pollinated seed set, number of seeds on the main shoot, number of pods on the main shoot, seeds per ten pods and plant height were measured for both the A2 and H2 populations and for a set of reference control cultivars. The H2 population had high segregation for pollen viability and meiotic stability, while the A2 population was characterised by low pollen fertility and a high level of chromosome loss. Both populations were taller, but had lower average fertility trait values than the control cultivar samples. The study also characterises fertility and meiotic chromosome behaviour in genotypes and progeny sets in heterozygous allotetraploid Brassica derived lines, and indicates that genotypes of the parents and H1 hybrids are affecting chromosome pairing and fertility phenotypes in the H2 population. The identification and characterisation of factors influencing stability in novel allohexaploid Brassica populations will assist in the development of this as a new crop species for food and agricultural benefit.