Impact of protein diversification on morphometric behavior of Andrographis paniculata Nees

Proteins are important biochemical parameters in genetic diversity and controlling morphological characteristics in plants. In this study, the proteomic and morphometric data of an important medicinal herb “Aci Paşa” (Andrographis paniculata) were combined together to illustrate their impacts on genetic variation of the plant’s population and to realize the connection between protein patterns and phenotypic behavior of the species. We used three protein extraction buffers including Tris, potassium phosphate, and sodium citrate. The Tris buffer was significantly different (p ≤ 0.01) than other two in terms of the quality and quantity of protein bands by producing 15 types of proteins ranged from 13 to 105 kDa of which two of them were polymorphic. Consequently, a total of 12 accessions of A. paniculata were subjected to morpho-proteomic analyses. The unweighted pair group method with arithmetic average cluster analysis of the accessions based on the protein data and morphological characteristics generated three and four clusters, respectively, at a Euclidean distance of 2.53 for the morphological traits. Moreover, seed proteins analysis revealed that the two polymorphic protein bands sized 20.5 (protein “b”) and 30 kDa (protein “a”) effectively diversified the morphological characteristics and phylogenetic relationships among the 12 accessions of A. paniculata. Interestingly, the protein “b” acted as an activator agent for the number of branches, leaves and total dry weight, while the protein “a” performed a suppressive role for the same traits. Additionally, the two high-weighted faint bands “c” (75 kDa) and “d” (100 kDa) with a very low expression in accession 11228 proved their suppressive role along with the “a” band, while these bands were strongly expressed in the rest of the accessions. These findings suggest that these four proteins should be sequenced and perfectly established for further proteomic analyses. Ultimately, the mentioned proteins can be developed for any prospective breeding program or gene identification

A rapid and effective method for dormancy breakage and germination of King of Bitters (Andrographis paniculata Nees.) seeds

Andrographis paniculata is a medicinal plant of the Acanthaceae family. The plant is important for its diterpenoids obtained from the mature plants grown normally through the seeds. However, the seed germination is slow and not efficient. Here, we report an effective method for breaking the seed dormancy to improve the germination per¬centage. Among the different physical and chemical scarification methods the fastest and the highest germination percentage were obtained from the seeds scarified with sandpaper. After three days of the scarification, 72.7% of the seeds germinated and this percentage was further increased to 90.4% after 15 days with a mean germina¬tion time (MGTa15) of 4 days. Although, the other physical and chemical treatments showed moderate effects on germination (GPa15 ranged from 7% to 64%), but they failed to reduce the MGTa15. Treating the sandpaper-scarified seeds with the seeds and the seeds coat protein extracts delayed the germination but did not affect the overall germination percentage, indicating the presence of unknown germinating factor(s) which may contribute to the dormancy of the seed. Together, the germination factor(s) and the hard seed coat caused the seed dormancy of A. paniculata which could be overcome by sandpaper scarification

Effects of different surface sterilizers on seed germination and contamination of king of bitters (Andrographis paniculata Nees).

Andrographis paniculata is a medicinal plant belongs to the plant family Acanthaceae. The plant is commonly propagated by seed. High contamination and low germination are two prevalent problems occur during germination and embryo growth stages. Approaches to decrease the seed contamination level and increasing germination percentage of Andrographis paniculata seeds were studied using a combination of chemical treatments with different sterilizer at different concentrations and exposure times in a factorial experiment based on Randomized Complete Block Design (RCBD) with three replications. Analysis of variance indicated that the effect of treatments were significant (p ≤ 0.01) on seed germination and contamination percentage (GP&CP), as well as on mean germination time (MGT), while non significant effects were observed on Number of Days to First Germination (NDFG) and Average Germination Percentage (AGP). The maximum (12.5%) and the minimum contamination percentage (2.2%) were observed in 4 weeks after treating in control treatment and treatment with 10% NaOCI for 10 min respectively. The results showed that 10% NaOCI for 10 min treatments is an effective option for decreasing the seed contamination percentage in this plant

Intraspecific Crossability in Andrographis paniculata Nees: A Barrier against Breeding of the Species

The ambiguity of crossability in Andrographis paniculata (AP) was pointed out in the present research. Accordingly, the effects of different style length and crossing time on intraspecific crossability of seven AP accessions in 21 possible combinations were investigated. The best results came out between 08:00 to 11:00 h for manual out-crossing of AP, while the time from 12:00 to 18:00 h showed a decreasing trend. Moreover, 12 mm style length was found as the most proper phenological stage in terms of stigmatic receptivity to perform out-crossing in this plant. All in all, AP behaved unlikely in each combination, and a significant difference was observed in crossability of AP accessions (P < 0.01). The lowest and highest crossability rate was found in hybrids 21 (11261NS × 11344K) and 27 (11322PA × 11350T) with 0.25% and 13.33%, respectively. Furthermore, a significant negative relationship between style length and crossibility (r2 = 0.762∗∗) was recorded in this research. As a final conclusion, crossing time and proper style length can improve the intraspecific crossability in the species, considerably. Despite all the mentioned contrivances, we still believe that a genetic incongruity should be involved as an additional obstacle in crossability of those combinations that failed or responded deficiently to outcrossing

Estimation of salt tolerance in Andrographis paniculata accessions using multiple regression model

The complexity and polygenic nature of the salt tolerance trait in plants needs to develop a multiple indicator in the screening process. The mentioned issue led us to carry out an experiment to identify tolerant genotypes through multiple parameters in Andrographis paniculata. For this purpose, the 40-days seedlings were grown in different salinity levels (control, 4, 8, 12 and 16 dS m−1) on Hoagland’s medium. The results indicated that salinity had a significant effect on the morphological, physiological and biochemical traits. All measured morphological traits, and chlorophyll, K+ and Ca2+ content were significantly decreased with increasing salinity levels, while proline and Na+ content increased. The present exploration revealed that, salt tolerance index (STI), using the multiple regression model, demonstrated a more stable trend than the single variable assay (total dry weight). Furthermore, STI based on multiple regression analysis gives an accurate definition of salt-tolerant individuals. Under salt stress, tolerant accessions had high STI and produced higher proline, K+ and Ca2+, and lower Na+ content than sensitive accessions. Cluster analysis based on related traits to STI, indicated high similarity in each group. These outcomes can be utilized to evaluate the salt tolerance threshold in the species and may have a great advantage over conventional methods. Probably, our upshots can be applied in the next breeding programs to develop salt-tolerant varieties

A critical review of the concept of transgenic plants: insights into pharmaceutical biotechnology and molecular farming

Using transgenic plants for the production of high-value recombinant proteins for industrial and clinical applications has become a promising alternative to using conventional bioproduction systems, such as bacteria, yeast, and cultured insect and animal cells. This novel system offers several advantages over conventional systems in terms of safety, scale, cost-effectiveness, and the ease of distribution and storage. Currently, plant systems are being utilised as recombinant bio-factories for the expression of various proteins, including potential vaccines and pharmaceuticals, through employing several adaptations of recombinant processes and utilizing the most suitable tools and strategies. The level of protein expression is a critical factor in plant molecular farming, and this level fluctuates according to the plant species and the organs involved. The production of recombinant native and engineered proteins is a complicated procedure that requires an inter- and multi-disciplinary effort involving a wide variety of scientific and technological disciplines, ranging from basic biotechnology, biochemistry, and cell biology to advanced production systems. This review considers important plant resources, affecting factors, and the recombinant-protein expression techniques relevant to the plant molecular farming process

A classical genetic solution to enhance the biosynthesis of anticancer phytochemicals in Andrographis paniculata nees

Andrographolides, the diterpene lactones, are major bioactive phytochemicals which could be found in different parts of the medicinal herb Andrographis paniculata. A number of such compounds namely andrographolide (AG), neoandrographolide (NAG), and 14-deoxy-11,12-didehydroandrographolide (DDAG) have already attracted a great deal of attention due to their potential therapeutic effects in hard-to-treat diseases such as cancers and HIV. Recently, they have also been considered as substrates for the discovery of novel pharmaceutical compounds. Nevertheless, there is still a huge gap in knowledge on the genetic pattern of the biosynthesis of these bioactive compounds. Hence, the present study aimed to investigate the genetic mechanisms controlling the biosynthesis of these phytochemicals using a diallel analysis. The high performance liquid chromatography analysis of the three andrographolides in 210 F1 progenies confirmed that the biosynthesis of these andrographolides was considerably increased via intraspecific hybridization. The results revealed high, moderate and low heterosis for DDAG, AG and NAG, respectively. Furthermore, the preponderance of non-additive gene actions was affirmed in the enhancement of the three andrographolides contents. The consequence of this type of gene action was the occurrence of high broad-sense and low narrow-sense heritabilities for the above mentioned andrographolides. The prevalence of non-additive gene action suggests the suitability of heterosis breeding and hybrid seed production as a preferred option to produce new plant varieties with higher andrographolide contents using the wild accessions of A. paniculata. Moreover, from an evolutionary point of view, the occurrence of population bottlenecks in the Malaysian accessions of A. paniculata was unveiled by observing a low level of additive genetic variance (VA) for all the andrographolides

Microsatellite-based evidences of genetic bottlenecks in the cryptic species “Andrographis paniculata Nees”: a potential anticancer agent

Andrographis paniculata (AP) is a medicinal plant species introduced into Malaysia. To address the genetic structure and evolutionary connectedness of the Malaysian AP with the Indian AP, a DNA sequence analysis was conducted based on 24 microsatellite markers. Out of the 24 primer sets, seven novel microsatellite primers were designed and amplified intra-specifically according to the available Indian AP sequences at the National Centre for Biotechnology Information (NCBI), where 17 of them were amplified using the cross-species strategy by employing the primers belonging to Acanthus ilicifolius Linn (Acanthaceae) and Lumnitzera racemosa Wild (Combretaceae). The primers were then applied on the Malaysian AP accessions. Sixteen of the new microsatellite loci were amplified successfully. Analysis of these microsatellite sequences, revealed some significant differences between the Indian and Malaysian AP accessions in terms of the size and type of the repeat motifs. These findings depicted the cryptic feature of this species. Despite identifying several heterozygous alleles no polymorphism was observed in the detected loci of the selected accessions. This situation was in concordance with the presence of “fixed heterozygosity” phenomenon in the mentioned loci. Accordingly, this was fully consistent with the occurrence of the genetic bottleneck and founder effect within Malaysian AP population. Apart from the amplification of new microsatellites in this species, our observations could be in agreement with the risk of genetic depletion and consequently extinction of this precious herb in Malaysia. This issue should be taken into consideration in the future studies

Genoproteomics-assisted improvement of Andrographis paniculata: toward a promising molecular and conventional breeding platform for autogamous plants affecting the pharmaceutical industry

Andrographis paniculata (Burm. f.) Wall. ex Nees. (AP) is a hermaphroditic, self-compatible, and habitual inbreeding plant. Its main bioactive component is andrographolide, which is capable of inducing autophagic cell death in some human cancer cells and helps fight HIV/AIDS. Increasing the andrographolide content by investigating the genetic mechanisms controlling its biosynthesis in order to improve and develop high-yielding cultivars are the main breeding targets for AP. However, there might exist some limitations or barriers for crossability within AP accessions. Recently, this problem was addressed in AP by using a combination of crossbreeding and biotechnology-aided genetic methods. This review emphasizes that development of a breeding platform in a hard-to-breed plant, such as AP, requires the involvement of a broad range of methods from classical genetics to molecular breeding. To this end, a phenological stage (for example, flowering and stigma development) can be simplified to a quantitative morphological trait (for example, bud or stigma length) to be used as an index to express the highest level of receptivity in order to manage outcrossing. The outcomes of the basic crossability research can be then employed in diallel mating and crossbreeding. This review explains how genomic data could produce useful information regarding genetic distance and its influence on the crossability of AP accessions. Our review indicates that co-dominant DNA markers, such as microsatellites, are also capable of resolving the evolutionary pathway and cryptic features of plant populations and such information can be used to select the best breeding strategy. This review also highlights the importance of proteomic analysis as a breeding tool. In this regard, protein diversification, as well as the impact of normal and stress-responsive proteins on morphometric and physiological behaviors, could be used in breeding programs. These findings have immense potential for improving plant production and, therefore, can be regarded as prospective breeding platforms for medicinal plants that have an autogamous mode of reproduction. Finally, this review suggests that novel site-directed genome editing approaches such as TALENs (Transcription Activator-Like Effector Nucleases) and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein-9 nuclease) systems together with other new plant breeding technologies (NPBT) should simultaneously be taken into consideration for improvement of pharmaceutical plants