Large Number Of Shoots Research Articles

Adoption on Cassava Technology Package to Improve the Plant Growth in Kalasan, Yogyakarta, Indonesia

Production of cassava in Kalasan is still low. The problem faced is to improve cassava growth, thereby increasing cassava yields. The aim of this study was to determine the effect of mycorrhizal inoculation, stem wounding soaked in PGPR, and spraying nano EPOFB ash fertilizer on the growth of cassava plants. The demonstration plot for planting cassava in Kalasan field was arranged in a completely randomized block design (CRBD), with single factor treatments, namely: A = stem without wounding (control), B = stem with wounding and soaked in PGPR, C = stem without wounding and inoculated with mycorrhiza, D = stem without wounding and sprayed with nano EPOFB ash fertilizer, E = stem with wounding + soaked in PGPR + inoculated with mycorrhiza + sprayed with nano EPOFB ash fertilizer. The results showed that the application of the innovative technology package (stem with wounding + soaked in PGPR + inoculated with mycorrhiza + sprayed with nano EPOFB ash fertilizer) to cassava cultivation in Kalasan had an effect on the percentage of mycorrhizal colonization, increased the dry weight of the roots, and accelerated the emergence of large numbers of shoots, thereby producing the highest number of leaves despite the shortest plant height.

In Vitro Shoot Culture Growth of Pontianak Orange (Citrus nobilis var. microcarpa) with the Addition of Naphthalene Acetic Acid and 6-Benzyl Amino Purine

Pontianak siam orange (Citrus nobilis var. microcarpa) is an important agricultural commodity unique to Sambas Regency, West Kalimantan. The demand for Pontianak siam oranges has been steadily increasing over the years, yet the production of Pontianak oranges has not been able to meet the domestic demand. The limited availability of superior seedlings is one of the reasons for this situation. Therefore, efforts are needed to fulfill the demand for Pontianak siam oranges by producing a large quantity of superior seedlings in a short period. One of the strategies that can be employed is in vitro propagation. This research aims to observe the growth of Pontianak Siamese orange shoot explants after administration of NAA (Naphthalene Acetic Acid) and BAP (6-Benzyl Amino Purine). This research was conducted from January to June 2023 at the Tissue Culture Laboratory of the Department of Biology, Faculty of Mathematics and Natural Sciences, Tanjungpura University. The study employed a Completely Randomized Design (CRD) with two treatment factors namely NAA concentrations with 4 levels (0; 0.25; 0.5; 0.75 mg/L), and BAP concentrations with 5 levels (0; 0.5; 1; 1.5; 2 mg/L). Parameters observed were the emergence time of shoot, shoot number, height of shoot, and the number of leaves. The results showed that single NAA, single BAP, and combination treatment of NAA and BAP significantly affected all growth parameters. Treatment with the addition of 0.25 mg/L NAA and 1 mg/L BAP (A1B2) was the treatment with the fastest shoot emergence time (2.80 days after planting), the largest number of shoots (2.60 shoots)the tallest shoots (1.50 cm), , and the greatest number of leaves (2.60 leaves).

In vitro regeneration of black pepper (Piper nigrum L.)

This investigation was done to determine how various plant growth regulators such as benzyl adenine (BA) and indole-3-butyric acid (IBA) affected the in vitro plant regeneration of the black pepper plant. The largest number of shoots was produced when the shoot tips and nodal segments of black pepper were utilized as explants and inoculated in Murashige Skoog (MS) media supplemented with 1.5 mg/L BA. The treatment with 2.0 mg/L BA at 8 weeks after induction (WAI) produced the most leaves (3.2). In the 2.0 mg/L treatment of IBA, the greatest number of roots was regenerated. The treatment 2.0 mg/L BA + 2.0 mg/L IBA produced the highest results for shoot induction, shoot length, and overall leaf production when the effects of both hormones were combined. The same treatment also produced the highest proportion of root induction and longest roots. Regenerated plantlets survive at a 46.67% rate in a shaded building and at a 57.14% rate in the open air under bright sunlight. In order to produce black pepper on a big scale, an effective approach for in vitro regeneration of black pepper has been established. Bangladesh J. Agri. 2023, 48(2): 75-85

Application of NAA and BA to Calotropis gigantea (L.) W.T. Aiton in vitro

Crown flower (Calotropis gigantea (L.)) is a weed plant potentially to be used as raw material for textile fibers yet the utilization is not optimal. Efforts to optimize the utilization of C. gigantea are needed. This research aimed to determine the appropriate concentration of plant growth regulators, including NAA (Naphthalene Acetic Acid) and BA (Benzyl Adenine) to stimulate optimal growth of C. gigantea to be used as the basis for in vitro propagation of C. gigantea. The research was arranged in a completely randomized design (CRD). Explants were planted on Murashige and Skoog (MS) medium with various concentrations of NAA (0; 0.5; 1; 1.5; and 2 ppm) and BA (0; 1; 2; 3; and 4 ppm). Based on the research result, the addition of 3 ppm BA followed by increasing the concentration of NAA to 1.5 ppm could increase the number of C. gigantea shoots, and giving 1 ppm BA followed by adding 1.5 to 2 ppm NAA increased shoot height. The best plantlet response was found in media with 0 ppm NAA and 4 ppm BA, which could produce an optimal shoot height of 5.9 cm and a large number of shoots of 5.67 shoots. Increasing NAA concentration retarded root formation and reduced the root length and number of leaves, while the medium without NAA gave the earliest root emergence of 11 days after planting, the number of leaves of 20 strands, and the longest root of 10.9 cm. Applying a single substance BA did not accelerate all the variables observed.

In vitro propagation of oil-bearing Rosa damascena using phloroglucinol: A protocol for rapid and high-quality shoot multiplication and rooting

Damask rose (Rosa damascena Mill.), of the family Rosaceae, is an important ornamental-medicinal, essential oil-bearing species. Due to its economic value, the aim of this study was to establish a new protocol for the in vitro propagation of R. damascena ‘Isfahan' using phloroglucinol. Axillary bud explants, Murashige and Skoog (MS) medium, and combinations of various plant growth regulators (PGRs) were used for the development of an effective method for rapid and high-quality shoot multiplication and in vitro rooting of Damask rose ‘Isfahan’. In the establishment step, 0.1 mg·L–1 phloroglucinol (PG), 0.2 mg·L–1 gibberellic acid (GA3), 0.1 mg·L–1 indole-3-butyric acid (IBA) and 1 mg·L–1 6-benzylaminopurine (BAP) were used. Secondary explants (nodal segments containing an axillary bud), obtained after 30 days of culture, were transferred to the proliferation media enriched with different concentrations of PG (0, 0.1, 0.2 and 0.3 mg·L–1), BAP (0, 1, 2 and 3 mg·L–1) and IBA (0, 0.1, 0.2 and 0.3 mg·L–1). A total of 64 combinations of these PGRs were investigated as a factorial experiment on the basis of a completely randomized design. Results showed that the shortest time until bud break (4.3 days), the highest number of green leaves (13.3) and lowest number of yellow leaves (1.3) were obtained in the explants grown on the medium containing 0.3 mg·L–1 PG + 2 mg·L–1 BAP + 0.2 mg·L–1 IBA. The largest number of shoots (4.8), and the highest fresh (5.9 g) and dry (1.2 g) weight of roots per explant were produced in the medium supplemented with 0.2 mg·L–1 PG + 2 mg·L–1 BAP + 0.3 mg·L–1 IBA. The highest regeneration coefficient (2.53), stem length (6.47 cm), fresh (2.32 g) and dry (0.51 g) weight of stem were obtained in the medium with 0.2 mg·L–1 PG + 2 mg·L–1 BAP + 0.2 mg·L–1 IBA. On the other hand, the highest number of roots (4.7) per explant were produced in the media supplemented with 0.3 mg·L–1 PG + 2 mg·L–1 BAP + 0.3 mg·L–1 IBA. None of the shoots regenerated roots in the absence of IBA. In most measured traits, the lowest values of the measured traits were obtained in the control medium without PGRs. Rooted plantlets were transferred to pots containing perlite and peat moss in 2:1 proportion and acclimatized in a greenhouse with a mean 82% survival rate. The positive role of PG in micropropagation was confirmed. The obtained results are more efficient than the previously established ones. Therefore, this cost-effective protocol can be used for the in vitro mass propagation of R. damascena Mill.

Effect of Different Types of Medium and Fertilizer on Acclimatization of Vanda tricolor

Vanda tricolor is an orchid endemic to Mount Merapi whose existence is threatened by the eruption of Mount Merapi which can occur at any time and the community has taken it directly from nature. Several attempts have been made to preserve the Vanda tricolor orchid, one of which is through in vitro culture. In vitro culture studies of Vanda tricolor have been carried out and large numbers of shoots have been obtained through multiplication and then raised by subculture. Plantlets obtained must be acclimatized to adapt and be returned to their original habitat. Growing medium and fertilizer application are important factors in the acclimatization process. This study aims to determine the best type of medium and fertilizer for acclimatization of Vanda tricolor orchids. This research was conducted using an experimental method in the preparation of a Factorial Completely Randomized Block Design. The first factor is the type of medium, namely coconut husk, moss and coconut husk + moss, the second factor is the type of fertilizer, namely without fertilizer, foliar fertilizer and liquid organic fertilizer. From the two treatment factors, 9 treatment combinations were obtained. Each treatment was repeated 3 times and each replication consisted of 3 samples. The results showed that there was a mutual influence between the type of medium and fertilizer. The coconut husk + moss is the best medium for acclimatization of Vanda tricolor orchids. Meanwhile, foliar fertilizer and liquid organic fertilizer compared to without fertilization, can increase the growth of Vanda tricolor orchids during acclimatization.

Propagation of Grape by Nodal Stem cuttings

Experiments were conducted to test the effect of MS saltsconcentration, potassium dihydrogen phosphate and growth regulators onshoot proliferation and growth in grapevine (Vitis vinifera L.), usingexplants derived from in vitro grown plantlets. Murashige and Skoogoriginal medium was used as basal medium. Shoot proliferation occurredprincipally when using nodal stem segments as explants. Among MS-saltstrengths tested, full MS- strength (1X) was optimum for shootproliferation and elongation. Increasing the original phosphorusconcentration in the medium by the addition of 170 mg/L KH2 PO 4 had apositive, marked effect on shoot number and shoot length. The nodal stemsegment explants responded best to relatively low concentrations of NAAand kinetin, and a large number of shoots and good plantlets height wereobtained on the medium containing 0.1mg/L NAA and 1.0 mg/L kinetin.Root formation occurred on the medium containing 0.3 mg/L IBA.

Effect of Explant Origin and Medium Plant Growth Regulators on In Vitro Shoot Proliferation and Rooting of Salvia tomentosa, a Native Sage of the Northeastern Mediterranean Basin

Salvia tomentosa is a perennial sage of the maquis vegetation, strongly aromatic, with medicinal and ornamental value. Aiming to facilitate the promotion of the species in the floriculture sector, its micropropagation was studied in terms of the effect of explant origin and medium plant growth regulators on in vitro cultures establishment, shoot proliferation and rooting. Shoot tip or single node explants excised from either in vitro grown seedlings or greenhouse plants grown from cuttings of mature plants were cultured on MS medium supplemented with 6-benzyladenine (BA) at concentrations 0.0–3.2 mg L−1 in combination with 0.0, 0.01 and 0.1 mg L−1 1-naphthaleneacetic acid (NAA). Infections were the biggest problem for establishing cultures from greenhouse plants, while explants from in vitro seedlings showed hyperhydricity, especially the nodal ones. In the multiplication stage, high BA concentrations reduced explant response to produce shoots as well as the length of shoots produced, and although they increased shoot number per responding explant, a large number of shoots were hyperhydrated especially in explants of seedling origin. In general, the highest multiplication indexes, relevant to normal shoots, were given by the hormone-free medium or one with low BA concentration, and by shoot tip explants originated from greenhouse adult plants. The highest rooting rates (70–78%) were achieved either at full or half strength hormone-free or with 0.1 mg L−1 indole-3-butyric acid (IBA) medium. Increasing IBA resulted in an increase in root number, but a decrease in rooting rate and root length. Ex vitro acclimatization was highly successful (92–96%) in peat-perlite (1:1 v/v) substrate. The micropropagation protocol developed will contribute to the promotion of S. tomentosa in the floriculture and pharmaceutical industry and will be a tool for breeding programs targeting high value products.

Molecular Assessment of Established Clonal Propagated Mulberry (Morus nigra L.)

Mulberry (Morus Sp.) is one of the economically important trees cultivated for the tasteful fruits, its several pharmaceutically important chemicals potential, timber, cosmetic and in silk industry for its foliage, also in various molecular breeding applications. The origins of most cultivated mulberry varieties are believed to be in the area of China-Japan and in the Himalaya foothills. and it now has a very wide distribution range in Asia-Europe (from Korea to Spain, including China, India, Central Asia, and Near East); in Africa (North and East Africa) and in America (from the U.S.A. to Argentina, including Mexico, Central America, Colombia and Brazil). In Saudi Arabia, Mulberry grown well and spread in different places such as Taif Province (El Shafa region), Eastern region; Al-Ehsaa Province and some southern areas. Nodal explants of Morus nigra were clonally propagated in vitro for plant regeneration. Auxiliary shoot buds have been promoted in Murashige and Scoog (MS) media in a variety of cultural contexts. The largest number of shoots (13.00 ± 0.47) with an average length of 2.00 ± 0.47 cm was initially obtained from a medium containing 2.0 mg / L N6-benzyladin (BA) and 3% sucrose. Recurrent subcultures provided the highest number of seedlings (approximately 29.30) for excavation after the fourth passage. Seedlings were rooted in 1/2 MS medium supplemented with 1.0 mg / 1 indole-3-butyric acid (IBA). Successfully, about 90% of the plantlets acclimatized. Along with the determination of the genetic variations between three mulberry genotypes including two cultivated accessions (Morus alba) and one wild genotype (M. nigra) utilizing inter-simple sequence repeat (ISSR) markers. Genetic variation and phylogenetic relationship of mulberry germplasm collection have been studied. All ISSR markers used in this study revealed higher genetic diversity in the wild species comparing with cultivated species. ISSR matrices reported that the mean genetic similarity coefficient was 0.7677 for all mulberry genotypes. Although some differences have been observed, much similarity has been obtained in dendrogram topology. Cluster analysis of the ISSR using UPGMA software revealed that wild species were genetically distinct. The correlation coefficients of similarity for the ISSR used are statistically important. The Principal Coordinate Analysis (PCA) for ISSR data also supports its UPGMA clustering. The average number of genetic variations recorded in mulberry genotypes was 0.287±0.096. Dendrogram (Unweighted peer group method analysis) classifies mulberry accessions into two main groups; Admissions collected from the western area of Taif, and the other comprised two sub-clusters including one isolate, i.e., M nigra, a collection from Al shafa. Contains access to another sub-cluster southwest regions of Taif, which belong to Morus nigra wild growing. These accessions of mulberry were found to be genetically similar to north and southwest Taif Province. These results have significant implications for improving the mulberry germ plasma characterization, conservation and investigates of the genetic diversity among the mulberry species grown in Taif governorate and to establish a micro-propagation system as germplasm conservation to preserve the assets of local mulberry and thus develops an easy and effective method to identify native genotypes in a limited space and time frame.

Can Artificial Intelligence Help in the Study of Vegetative Growth Patterns from Herbarium Collections? An Evaluation of the Tropical Flora of the French Guiana Forest.

A better knowledge of tree vegetative growth phenology and its relationship to environmental variables is crucial to understanding forest growth dynamics and how climate change may affect it. Less studied than reproductive structures, vegetative growth phenology focuses primarily on the analysis of growing shoots, from buds to leaf fall. In temperate regions, low winter temperatures impose a cessation of vegetative growth shoots and lead to a well-known annual growth cycle pattern for most species. The humid tropics, on the other hand, have less seasonality and contain many more tree species, leading to a diversity of patterns that is still poorly known and understood. The work in this study aims to advance knowledge in this area, focusing specifically on herbarium scans, as herbariums offer the promise of tracking phenology over long periods of time. However, such a study requires a large number of shoots to be able to draw statistically relevant conclusions. We propose to investigate the extent to which the use of deep learning can help detect and type-classify these relatively rare vegetative structures in herbarium collections. Our results demonstrate the relevance of using herbarium data in vegetative phenology research as well as the potential of deep learning approaches for growing shoot detection.

Micropropagation of pokeweed (Phytolacca americana L.) and comparison of phenolic, flavonoid content, and antioxidant activity between pokeweed callus and other parts.

BackgroundPokeweed (Phytolacca americana L.) is regarded as an invasive plant in many parts of the world but possesses therapeutic characteristics used for antitumor and rheumatism treatment. This study investigated the effects of auxins and four explants on pokeweed callus induction. The effects of cytokinins and combinations between cytokinins and NAA on shoot and root induction were also studied. TPC, TFC and antioxidant activity of calli were screened and compared with other pokeweed plant parts.MethodsFour explants were used to induce callus using 2,4-D and IBA at 1, 2, 3 and 4 mg/l for each auxin. Direct shoot organogenesis from nodal explants was investigated using BAP, kinetin and TDZ (1, 2 and 4 mg/l for each cytokinin). Combined effects between cytokinins and NAA at 0.1, 0.2 and 0.3 mg/l were further simultaneously estimated with root induction. Calli derived from the leaves were compared with other plant parts for TPC, TFC and antioxidant activity using the Folin-Ciocalteu, AlCl3 colorimetric assay and DPPH assays, respectively.ResultsResults showed that MS medium containing 2 mg/l 2,4-D induced callus formation on leaf explants that provided highest fresh and dry weights. Three types of synthetic cytokinins as kinetin, TDZ and BAP were used for direct shoot organogenesis from pokeweed nodes. MS medium containing 2 mg/l kinetin was effective in stimulating normal shoots, with the largest number of shoots and leaves and the longest shoots. The combination between cytokinins and NAA showed no positive effect on shoot and root induction from pokeweed nodal explants. For TPC and TFC determination, pokeweed seeds and leaves possessed the highest phenolic and flavonoid contents, respectively. Highest phenolic content of pokeweed seeds led to lowest IC50 by DPPH assay. Phenolic content was higher than flavonoid content.ConclusionResults suggested promising conditions for callus induction. Leaf explants cultured on MS medium with 2 mg/l 2,4-D and nodal explants cultured on MS medium with 2 mg/l kinetin provided the largest number of normal shoots and leaves. NAA did not show positive effects on shoot and root induction when combined with cytokinins. Chemical constituent screening indicated that seeds and leaves provided highest TPC and TFC, respectively, while pokeweed calli contained higher phenolic than flavonoid content. This is the first report describing chemical constituent screening and antioxidant activity of calli and other parts of the pokeweed plant. Results provided significant information to further enhance bioactive compound contents of pokeweed calli using elicitation methods.

ЦИКЛ «ВВЕДЕНИЕ В КУЛЬТУРУ IN VITRO – МИКРОРАЗМНОЖЕНИЕ» У СОРТОВ ВИНОГРАДА: ЛЕДЯНОЙ, ЗОЛОТЦЕ, МУСКАТ АКСАЙСКИЙ

The article notes the necessity to improve clonal micro-propagation in order to create genetic banks. The key moment is to optimize the "introduction to in vitro culture- micro-propagation" cycle. The possibility of joint use of apical meristem and chemotherapy was studied for three grapevine varieties that differ in morphobiological indicators. To do this, the antiviral drug Ribavirin, salicylic ac-id, the antibiotic Cefotaxim, the growth regu-lator Melafen, were supplemented to the nu-trient medium. A detailed description of the research methods, both generally accepted and improved by the author are given in the paper. Intensive development of apical meri-stem in Ledyanoy variety has been detected, and the meristem of the Zolotce variety has been found to have a positive reaction to all the drugs studied, especially Cefotaxim and Melafen. The Muscat Aksaiskiy had signifi-cant death and slow development of apical meristem. In variants with the use of Ribavi-rin, Salicylic acid and Cefotaxima a better development of meristems was marked. There were more large meristems and with the signs of growth. The positive effect of chemotherapy on the multiplication coeffi-cient and on the shoot formation was estab-lished. The reproduction rate of Ledyanoy variety increased by 1.8 — 2.7 times, the Zolotce variety by 2-2.3 times, and the Mus-cat Aksayiskiy variety by 2.3-2.6 times. The largest number of shoots was formed in the varieties Ledyanoy and Muscat Aksayskiy - 53 pcs., slightly less in the variety Zolotce -47 pcs. In the Ledyanoy variety, the best options are: Cefotaxime and salicylic acid; in the Zolotce variety, Cefotaksim; in the Muscat Aksaiskiy variety: Cefotaxime and salicylic acid. The necessity of using chemotherapy at the stage of introducing apical meristems into the culture in vitro to optimize clonal micro-reproduction was proved.

In vitro plant regeneration of Christmas cactus (Schlumbergera truncata (Haw.) Moran) by indirect morphogenesis

Abstract Plants of Schlumbergera truncata (Haw.) Moran were obtained by indirect morphogenesis from the segment section of shoots in vitro, they were multiplied and rooted. Also were determined the effect of the lighting regime, the composition of the nutrient medium on the consistency and frequency of callus formation. The studies were conducted during 2016–2018. The mode of effective sterilization (more than 90%) of S. truncata plant explants using 0.1% HgCl2 for 7–8 min was established. Optimal conditions for the induction of callus formation in stem node segments of S. truncata plants (rate more than 90% and significant growth) were created on MS (Murashige and Skoog 1962) nutrient medium supplemented with 1.0 mg/l BAP (6-benzylaminopurine) and 0.3 mg/l NAA (1-naphthylacetic acid) under conditions of placement on the nutrient medium and doing a significant number of cuts on the explants. The light intensity of 2.0–3.0 klx, obtained by a callus of dense consistency of dark green pigmentation, when using the thermostat condition without illumination, the callus had loose consistency, dark yellow pigmentation. It is established that the influence of the lighting regime and the composition of the nutrient medium on the frequency of callus formation is statistically significant. The largest number of shoots was obtained on the MS medium with the addition of 2.0 mg/l of BA. At the same times, shoot proliferation and root induction in such numbers were observed on MS culture medium with the addition of 0.5 mg/l BA and 0.5 mg/l kinetin (multiplication factor – 8.8±0.6 per 60-day cultivation cycle).

A protocol for fast in vitro propagation of Impatiens hawkeri cv. Sweeties Blue Star, an important ornamental-cover species

Abstract Impatiens hawkeri is among the three important bedding plants in the world. There is no efficient protocol for fast micropropagation of Impatiens hawkeri cv. Sweeties Blue Star. Single nodes were germinated on Murashige and Skoog (MS) medium supplemented with 1-naphthaleneacetic acid (NAA; 0.00, 0.10 and 0.50 mg l-1), 6-benzyladenine (BA; 0.00, 0.10, 0.50 and 1.00 mg l-1) and kinetin (KIN; 0.00, 0.10, 0.50 and 1.00 mg l-1). The shoot length was highest in medium containing 0.10 mg l-1 NAA and control. The largest number of shoots (14.06 and 14.00 per explant) was produced in media supplemented with 1.00 mg l-1 BA, 0.50 mg l-1 NAA and 1.00 mg l-1 BA, 0.10 mg l-1 NAA, respectively. The 0.10 mg l-1 NAA along with 1.00 mg l-1 BA was found to be superior for production of leaf (57.13). Maximum root length (33.80 mm) and root number (29.13) were obtained on medium supplemented with 0.10 mg l-1 NAA without BA and KIN. Plantlets were transplanted to pots filled with perlite and peat moss (in ratio of 1:1) and transferred to the greenhouse for acclimatization. Regenerated plantlets were morphologically identical with mother plants. Upon ex vitro transfer, 100% of plants survived.

Diodos emissores de luz (LEDS) no cultivo in vitro de Eugenia involucrata

Esse trabalho objetivou avaliar o efeito do tipo de luz no cultivo in vitro de Eugenia involucrata. Os tratamentos consistiram do cultivo sob diodos emissores de luz (LEDs) de cor vermelho, azul, verde ou branco e lâmpadas fluorescentes. Nos cultivos sob LEDs branco e azul foram observadas as maiores médias de estabelecimento; sob o azul e lâmpadas fluorescentes, o maior número de folhas; sob o azul, o maior número de brotos; sob o branco, menor contaminação bacteriana; e sob o verde os piores resultados. LEDs azul, vermelho, branco e lâmpadas fluorescentes influenciam favoravelmente a propagação in vitro de E. involucrata.

К БИОЛОГИИ И ЭКОЛОГИИ ГОРЧАКА ПОЛЗУЧЕГО (ACROPTILON REPENS) НА ЮЖНОМ УРАЛЕ

The article provides information on the biology of Acroptilon repens (L.) DC. This species is a perennial root weed from the Asteraceae family. It is included in the list of quarantine plants of the Russian Federation. In the South Urals the species appeared not long ago and is classified as potentially invasive. In the Republic of Bashkortostan the first record of the species dates back to 2013. The aim of the research was to study the biological characteristics and parameters of coenopopulations (CP) of a dangerous quarantine plant under new living conditions in the Republic of Bashkortostan in comparison with the previously formed invasive coenopopulations of the Orenburg region. Three localities of invasion of A. repens were examined (1 in the Republic of Bashkortostan and 2 in the Orenburg region) during the expedition trips in 2018. The research specifies that the maximum height of plants (47.9 cm) and most other morphometric parameters are registered in the Shkunovka CP. The highest percentage of the species participation in the community is also observed there (82.9 %). The largest number of shoots per 1 m2 (31.3) was found in the Kumertau CP. The Divnopolye CP and Shkunovka CP have similar indexes of morphometric parameters. The Kumertau CP is different from the CPs of Orenburg region in most parameters. Predominance of individuals of the highest class is noted in two coenopopulations of Orenburg region. They are classified as prosperous coenopopulations and Kumertau CP as a depressive one. The discriminant analysis proves that in all coenopopulations of A. repens, individuals are morphostructurally different, with no overlap between them. The Kumertau CP is phenotypically the most distant from the Divnopolye CP and Shkunovka CP.

Detection and identification of hybrids in the coenopopulation of co-growing species of Cypripedium (Orchidaceae) in the Novosibirsk Region

Cypripedium calceolus L. and C. macranthon Sw. (Orchidaceae) are rare species throughout almost the entire territory of Russia. Though the data on intraspecific genetic differentiation has been presented in the literature, the species of C. calceolus, C. macranthon and alleged hybrids of C. × ventricosum growing in sympatric population in the Iskitim District of the Novosibirsk Region (NR) have not been investigated yet. The purpose of this work is dentification of the rare species of Cypripedium calceolus, C. macranthon and detection of C. × ventricosum hybrids in the NR using morphometric parameters and ISSR markers. It was found the mature generative specimens of C. × ventricosum formed the largest number of shoots, compared with the parent species. According to the number of flowers on the generative shoot, the hybrid occupied an intermediate position. According to the ISSR analysis, components from both parental species were identified in each of the three hybrids studied. Therefore, the presence of C. × ventricosum hybrid plants in the local NR coenopopulation of co-growing species C. calceolus and C. macranthon was detected and confirmed based on the study of morphometric parameters and ISSR markers.

PERBANYAKAN TANAMAN HIAS AIR TAWAR Alternanthera reinickii DAN Ludwigia sp SECARA IN VITRO GUNA MENDUKUNG AQUASCAPE

Water plants are the hydrophytic plants that occupy the highest ecology in aquatic environments. 
 Alternanthera reinickii and Ludwigia sp are freshwater ornamental plants that are in great deman d of 
 aquascape lovers market. Plant tissue culture is a technique to grow the growing parts of plants, 
 either in the form of cells, tissues or organs in aseptic conditions in vitro. The study was conducted on 
 15 January to 15 April 2018, at the Center for Biotechnology Research and Bogor Agricultural 
 Genetic Resources. This research is done by using experiment method in laboratory. From the results 
 of research on ornamental plants Alternanthera reinickii data obtained on the leaf paremeter giving 
 regeneration substances grow the type of BAP with a concentration of 0.3 ml / l capable of producing 
 the largest number of leaves. In shoot height parameters obtained the highest results using Kinetin 
 treatment concentration 0.1 ml / l. In the parameter of shoot num ber of growth regulator substances 
 BAP concentration of 0.5 ml / l resulted in the largest number of shoots. In the parameter of the root 
 number of BAP treatment with the concentration of 0.3 ml / l shows the largest number of roots. In 
 contrast to the ornamental plant Ludwigia sp BAP treatment of 0.1 ml / l concentration is able to 
 produce the highest number of leaves In the parameters of shoot BAP treatment at low concentrations 
 of 0.1 ml / l. on the parameters of BAP treatment buds concentration of 0.3 m l / l resulted in the 
 largest number of shoots. In the parameter of the root number of BAP treatme nt, the concentration of 
 0.1 ml/l showed the highest number of roots.
 Keywords: Alternanthera reinickii, Ludwigia sp, Tissue culture, BAP, Kinetin

REGENERATION SYSTEM OF LETTUCE (<i>LACTUCA SATIVA</i> L.) THROUGH <i>IN VITRO</i> CULTURE

The article is devoted to the problem of obtaining regenerative plant varieties of lettuce (Lactuca sativaL.) Bouquet, Emerald, Chameleon (FSBSI Federal Scientific Vegetable Center). Salad culture is insensitive toin vitroconditions and requires the development of alternative regeneration conditions. Seeds were surface sterilized in 96% ethanol (30 s), then 15 minutes in 50% aqueous solution of "Whiteness" with the addition of Twin-20 (1 drop per 100 ml), washed three times in sterile distilled water. Then lettuce seeds were placed in Petri dishes on Gamborg B5 medium (2% sucrose, 3,0 g/l fitogeli). After 10 days, the obtained seedlings were cut, the leaf blades were cut into squares 1 cm in size and transferred to medium MS B5 (2% sucrose) with three variants of hormone concentrations: 0.1 mg/l NAA and 1 mg/l BAP, 1 mg/l NAA and 1 mg/l BAP, 1 mg/l NAA and 0.1 mg/l BAP. The formed shoots were rooted on MS medium (2% sucrose, 7,0 g/l agar). Cultivation was carried out on racks with fluorescent lamps at 25°C and a photoperiod of 14 hours, illumination of 2,5 thousand Lux. According to the results of studies, the concentration of hormones (0,1 mg/l NAA, 1 mg/l BAP) was selected, which allowed to obtain up to 10 shoots from one leaf explant. Moreover, the largest number of shoots was noted on leaf explants located closer to the veins of the sheet. The obtained data allowed to improve the system of regeneration of varieties of emerald lettuce, Bouquet and Chameleon.

Acceleration of Echinacea purpurea (L.) Moench Shoot Growth by Benzyl Adenine and Indole Butyric Acid Addition

Echinacea (Echinacea purpurea (L.) Moench) is a medicinal plant known to boost the immune system. Propagation is necessary to increase production. One of the methods of propagation in tissue culture. This research was conducted to understand the most suitable concentration of plant growth regulators. The treatment was given a combination of Benzyl Adenine (BA) and Indole Butyric Acid (IBA) with BA concentration of 1 ppm, 2 ppm, 3 ppm and 4 ppm while the IBA concentration used was 1 ppm and 2 ppm. The next step was subculture by using the combination among IBA 0 ppm, 0.5 ppm, 0.75 ppm and BAP 0 ppm, 0.5 ppm, 0.75 ppm of BAP. The result showed that the most shoots produced by the combination treatment of BA 2 ppm and IBA 1 ppm while the highest shoot and leaf number is best produced in treatment BA 1 ppm and IBA 2 ppm. The largest number of shoots was shown by treatment BA 2 ppm and IBA 1 ppm. This study can be concluded that BA 1 ppm and IBA 2 ppm, and BA 2 ppm and IBA 1 ppm gave the best treatment for shoot growth and control for root induction.