Microsatellite Marker Based DNA Fingerprinting of Cotton (Gossypium Spp.) Hybrids and Their Parents

Cotton production in India by vast majority comes from cotton hybrids whose genetic purity is of great significance in seed production chain and trade. Therefore, there is need to develop a rapid, reliable and reproducible technique to assess the genetic purity of cotton hybrids as traditional, morphological traits-based ‘Grow-Out Test’ is resource intensive, time consuming, tedious and not an infallible procedure. In this regard, a study was planned to understand the genetic diversity among the hybrids and their parents, and also to identify SSR markers for confirmation of genetic purity or hybridity. One intra-arboreum hybrid, CICR2 (DS 5 GMS × LD 327 Sel.), four intra-hirsutum hybrids viz., CSHH198 (CSH 19 × CSH 8), CSHH238 (SH 2379 9Y × PIL 8 Sel.), CSHH243 (CSH 2013 × CSH 43), CSHH1862 (GMS 16A × CB 33) and one hirsutum × barbadense hybrid, Phule 388 (RHC-006 × RHCb-001) along with their respective parental lines were selected for molecular characterization. Of the total 215 SSR markers surveyed, 60 markers conveyed polymorphism. The information conveyed by the polymorphic SSR markers was utilized to assess the molecular divergence among the study material. Maximum genetic dissimilarity of 0.66 was noted between Phule 388 and LD 327 (Sel.), and between RHC-006 and DS 5 (GMS). Minimum genetic dissimilarity of 0.07 was observed between CSHH1862 with CB 33, followed by 0.11 between CICR2 with DS 5 (GMS). SSR markers were highly efficient in capturing both intra-species and inter-species level diversity. The clustering and factorial analysis was in congruence with the species of Gossypium. The diploid species genotypes were clustered separately and distinctly from the rest of the genotypes. All the hirsutum hybrids and their respective parents were found closely clustered. The inter-specific hybrid, Phule 388 along with its parents was found grouped closely. The genetic purity of the hybrids was confirmed using identified SSR markers [GH486, BNL1421, BNL3594, JESPR151 for G. hirsutum hybrid CSHH198; GH486, BNL2449, JESPR151, TMB0436 for G. hirsutum hybrid CSHH238; BNL2449, JESPR151, JESPR152 for G. hirsutum hybrid CSHH243; and, GH527, BNL3812, TMB1484, TMB1645, NAU1190, BNL3816 for inter-specific G. hirsutum × G. barbadense hybrid Phule 388]. The SSR markers were efficient in analysis of hybrid seed purity. The information generated in the present study about genetic diversity and genetic purity testing will greatly facilitate quality seed production of these cotton hybrids and thus, better cotton production.

Varietal identification and fingerprinting of Pearl Millet (Pennisetum glaucum L.) varieties and hybrid using morphological descriptors and SSR markers

Pearl Millet (Pennisetum glaucum) is the sixth most important cereal crop in the world. The genomic resources available in Pearl millet can be utilized for fingerprinting and screening of hybrids using SSR markers and will be helpful for the assessment of seed purity. Hence, the present study was focused on fingerprint popular pearl millet varieties and hybrids of Tamil Nadu for varietal identification and hybrid purity test. The varieties used for DNA fingerprinting were CO (Cu) 9, CO 10, Pearl Millet hybrid CO 9 along with the parents, A' line ICMA 93111A and R' line PT 6029-30. The morphological features were recorded to screen the cultivars. The Pearl millet hybrid CO 9 scored the highest value for more than four quantitative characters via., Number of productive tillers (4-6), Leaf blade length (60-68cm), Leaf blade width (4.0-4.5cm), number of nodes (8-10), and 1000 seed weight (13-14g) which is at par and comparable with the composite CO 10 and higher than that of the variety CO (Cu) 9. PCR was performed using 36 SSR primers to find out polymorphism among the varieties. The SSR markers ICMP3021 and PSMP2089 were able to selectively identify CO (Cu) 9 from the other varieties. Whereas, the SSR markers ICMP3018, PSMP2219, and PSMP2220 were used to distinguish CO 10 from the other varieties. Further, the CO10 variety produced additional alleles for all the markers due to its composite nature. Among the thirty-six SSR primers screened, none of them were found suitable to distinguish the TNAU hybrid CO 9 from its parents. The unique DNA fingerprints developed in the present study can be utilized for seed purity testing and varietal identification.

Blend wheat AL-type hybrid and using SSRs to determine the purity of hybrid seeds

Heterosis is a promising approach to increase wheat yield from a limited planting area. In this study, a fine quality restorer line 99AR144-1 and three stable male sterile lines, AL18A, AL36A and AL20A, were assigned as male and female, respectively. Seeds of the wheat line 99AR144-1 and three male sterile lines were mixed according to different proportions and then planted at an experimental farm at the Xinjiang Academy of Agri-Reclamation Sciences from 2013 to 2016. When the mixed sowing ratios of combinations 2 (AL36A × 99AR144-1) and 3 (AL20A × 99AR144-1) were 6 to 8%, the seed production yields were higher than the control; the yield of hybrid seed production increased by 98.8 and 19.9%, respectively. This increase was attributed to a rise in the outcrossing seed setting rate. Further, this study used the Xbarc-8 SSR (simple sequence repeat) molecular marker to identify the purity of blend hybrid seeds and establish a regression equation for hybrid seed purity testing. The coefficient of the regression equation were 0.9878 and 0.9689 respectively, which shows that the purity of hybrids can be accurately predicted by using this equation. This method can quickly and accurately identify the seed purity in mixed seed production.

DNA fingerprinting of foxtail millet (Setaria italica L.) variety ATL 1 using SSR and RAPD markers along with morphological descriptors

Foxtail millet (Setaria italica) is a cultivated nutritional cereal, which originated in South Asia and is considered one of the oldest cultivated millets in India. DNA fingerprinting is mandatory for registration of newly developed varieties with National Bureau of Plant Genetic Resources (NBPGR) and Protection of Plant Varieties and Farmers' Rights Authority (PPV&FRA). Due to the limited availability of genomic information in foxtail millet, the use of DNA based markers in fingerprinting of crop varieties is also limited. Hence in the present investigation, available RAPD and SSR markers of cereals are used for fingerprinting the foxtail millet varieties. The newly released variety ATL 1 is differentiated from popular variety CO (Te) 7 using SSR and RAPD markers. About 66 maize SSR primers, 16 sorghum SSR primers, and 10 RAPD primers were used in the study. Out of 66 maize SSR markers used for study, one showed polymorphism. The marker umc1704 showed polymorphism between CO (Te) 7 and ATL 1 by the presence of 670 bp allele CO (Te) 7. The RAPD primers OPB4, OPA5, OPA11 and OPB1 also helped for differentiation of the two varieties. The identified makers will help for genetic purity testing of CO (Te) 7 and ATL 1 in the seed chain.

Seed genetic purity testing of F1 Benincasa hispida var. chieh-qua hybrids using SSR molecular marker analysis

To ensure that farmers can access high-quality seeds, it is essential to find a simple, rapid and accurate method to assess seed purity. In recent years, heterosis in chieh-qua has been widely applied to production. Using the whole genome sequence of chieh-qua, we designed simple sequence repeat (SSR) primers specific for chieh-qua. The parental lines of nine hybrids were screened using 200 SSR primers, seven of which exhibited polymorphisms. The bands were clear, stable and reproducible. We found dominant and co-dominant bands between the parents of the nine hybrids. The seven pairs of SSR primers were successfully used to assess genetic purity of the nine chieh-qua hybrids. The SSR molecular marker purity assessment results were consistent with the results obtained from a field grow-out test (GOT). However, the use of SSR markers provided a more accurate, reliable and faster method for seed purity testing than the GOT. We propose using SSR molecular marker technology to assess the genetic purity of chieh-qua hybrid seeds. With this method, the seed quality can be determined faster, which may help to accelerate the chieh-qua breeding process.

Spectrophotometrical purity and stability testing of a photosensitizer based on an indotricarbocyanine dye

The express method for testing purity and stability of an indotricarbocyanine dye based photosensitizer for photodynamic therapy was developed. Storage of the hydrophilic photosensitizer under unfavorable conditions resulted in contamination with a hydrophobic dye. The absorption spectrum of the hydrophobic dye was characterized by a new band at 514 nm that was absent in the absorption spectrum of the photosensitizer. There was no spectral overlap between this absorption band and the absorbance of the photosensitizer. Therefore, presence of the band at 514 nm allowed us to detect contamination of the photosensitizer substance with the hydrophobic dye. It was established that the optimal photosensitizer concentration for purity testing is ca. 0.8 mmol/L, when as little as 0.6 wt. % of the impurity can be detected. The developed method is not time consuming and requires accessible equipment only. Data obtained with this method was verified with the aid of liquid chromatography-mass spectrometry.

Diagnostic and Measurement System for Marine Engines

Modern way of machines’ exploitation, due to their high level of structural complication, requiresproper level of supervising. This is enabled by more than 60 years’ development of the inspection andmaintenance technologies. Nowadays it focuses on preventing the unplanned accidents by detection of prefailure states and predictive a future trend of the machine. To detect the problem and avoid the failure veryimportant and essential is to develop the Non Destructive Testing (NDT) and Condition Monitoring includingassessment of working process, vibration analysis, oil purity testing, endoscopic, ultrasonic and thermalanalysis. The paper presents a design and structure of the Exploitation Decision Aid System dedicated fordiagnosis of marine engines condition. There are also presented examples of results obtained with use of thesystem.

Evolution of liquid chromatography: Technologies and applications

Liquid chromatographic offers efficient analyte separation employing high pressure pumps. The reversed phase high performance liquid chromatography (RP-HPLC) is widely utilized in the purity testing and quantitative determination of pharmaceuticals and neutraceuticals. The limitations of traditional liquid chromatography such as particle size, resolution and selectivity demanded for the developments and Waters Corporation developed ultraperformance liquid chromatography (UPLC). Ultrafast liquid chromatography (UFLC) is another milestone, which offers faster and efficient separation. Multidimensional UHPLC provides separation of complex molecules. The particle size decrease enhances the resolution of LC separation. Ethylene bridged hybrid (BEH), Charged surface hybrid (CSH) and Peptide separation technology (PST) offer better performance in. The amalgamation of chromatographic and spectroscopic detectors namely fluorescence detector (FD) and mass spectrometry (MS) provides efficient separation. Liquid chromatography (LC) offers the analysis of pharmaceuticals, biological, food materials, and natural products. This review covers technologies and recent pharmaceutical and biomedical applications of liquid chromatography technologies