CC BY
20
УДК 578.08
DOI: 10.36305/0513-1634-2020-134-17-23
MICROMORPHOLOGY OF SEED IN SOME VICIA TAXA BELONGING TO SECTION
CRACCA FROM AZERBAIJAN
Kamala Valeh Asadova, Aydin Musa Asgarov
Genetic Resources Institute of ANAS, Azadlig Ave. 155, Baku, AZ1106, Azerbaijan;
E-mail: esedzadeo@gmail.com
For the first time, some taxa of the genus Vetch (Vicia L.) belonging to subgenus Vicilla and section Cracca (according to the system of Kupicha) from the Azerbaijan were subjected to morphological and scanning electron microscopic (SEM) analyses: V. tenuifolia Roth subsp. variabilis (Freyn & Sint. ex Freyn) Dinsm., V. villosa subsp. villosa Roth, V. villosa Roth subsp. varia (Host) Corb. (V. dasycarpa auct.), V. monantha Retz. (V. cinerea M. Bieb.), V. elegans Guss.
Seed patterns were collected in 2015-2017 from different region of Azerbaijan (Zagatala, Ordubad and Shamakhi). In all seed samples, the seed's surface was papillate. However, the size of the ribs in the papillas and the density of papillas were different in different taxa. The papillae with medium density were observed in V. tenuifolia subsp. variabilis. Dense papilla were observed in V. villosa subsp. villosa, V. villosa subsp. varia, V. monantha and V. elegans. V. villosa subsp. varia had conical, multirib papillas with waxy specks. However, small conical multirib were observed in V. tenuifolia subsp. variabilis and V. monantha. Conical multirib was found in V. villosa subsp. villosa and broad conical in V. elegans. Ovate hilum was observed in V. villosa subsp. varia. Oblong-elliptic hilum is peculiar to V. tenuifolia subsp. variabilis, V. villosa subsp. villosa, V. monantha, and V. elegans. The largest seeds are found in V. tenuifolia subsp. variabilis, and the smallest ones in V. villosa subsp. varia. Detected micromorphological features play an important role in the determination of different taxa of the genus Vicia L.
Keywords: seed, SEM, vetch, leguminous, intraspecific, section, species
Introduction
Azerbaijan is the biggest country of Southern Caucasis according to its area and population. The Republic that is situated in the junction of Europe and Asia has got a unique geographical position. Location in the subtropical latitude, richness and diversity of its nature are the factors that constantly influence the progress of Azerbaijan.
Azerbaijan is distinguished not only by climate and diversity of its soil types, but also by the genetic resoursces of this plants. Even thoug natural flora contains rich genefund of important food, technical, medicine, vitamin rich plants, only a part of this wealth is used [2].
Vetch (Vicia L.) is one of the most widespread genera of the family Fabaceae Lindl. As with other leguminous plants, taxa of Vicia are also selected for their nitrogen-fixing properties. In the roots of the plant, there are nitrogen-fixing bacteria that adopt the atmospheric free nitrogen and convert it into a mineral form that plants can use. During the vegetation period, vetches release these mineral substances into the soil through the roots that enriches the microflora of the soil and fertilize it. Due to this, the productivity of plants grown on the soil after the vegetation period and the quality of crop products rises [17]. Representatives of the genus Vicia L. are considered to be good predecessors. This genus contains 150-210 species occurring in Europe, Asia, North America, temperate regions of South America, and partly in tropical Africa [15; 25]. It has been established that in the flora of Azerbaijan the genus Vicia is represented by 2 subgenera, 11 sections, 39 species, 5 subspecies and 8 varieties ranging in their distribution from the sea level to mid-mountain zones [1; 3; 4].
The monographic study of the genus was carried out by F.K. Kupicha [23; 25; 26]. Using the classic methods, Kupicha, based on selected morphological signs, subdivided
Vicia into two subgenera: Vicia and Vicilla (Cracca). These subgenera differ in the relative length of the inflorescence and the presence or absence of nectar spots on the stipule. In their taxonomic studies of the genus Vicia, many scientists used the seed morphology [6; 11; 16; 24; 27; 28; 33], including micromorphology of the seed texture [7; 11; 12; 13; 16; 22; 29].
Signs of the seeds can be used as an important feature in the classification of species and in the determination of the phylogenetic relationships of intraspecific taxa.
In the first description of a number of vetch taxa, as well as in the Floras (Azerbaijan, the Caucasus flora), no signs of the seeds were mentioned. With this in mind, for the first time were studied the morphology and especially micromorphology of seed samples of species and subspecies of Vicia L. distributed in the flora of Azerbaijan.
The aim was to clarify the relationship between the intraspecific groups belonging to the genus Vicia L.
Material and methods
The material for the study consisted of seeds of Vetch, which we collected during the expeditions to different regions of Azerbaijan under the quidance of A. Asgarov in 2015-2017. Information on the collection area, biotope, geographical coordinates and altitude of each seed sample was indicated in the special descriptor forms (Table 1). We indicated information on the collection area, biotope, geographical coordinates and altitude of each seed sample in the special descriptions. Hipsometric altitude and area coordinates were measured with a Garmin eTex 20 model GPS device according to WGS-84 (G873) (World Geodetic System).
The most commonly accepted classification is the classification of Kupicha, so we accepted that classification in own work [23]. However, it should be noted that new phylogenetic results indicate that the genus Vicia, as traditionally understood, is not monophyletic [4; 9; 10; 32]. We used terminology adopted by some authors cited in our article [3; 10; 11; 14; 15; 18; 19; 20; 22; 24, 29].
In the samples, the surface structures, shape, density or waxy layer of papillae (buds with suction surface) were studied by scanning electron microscopy (SEM).
SEM analysis of plant seeds has been done on a microscope JEOL JSM 6610 LV. Modern plant seeds (both living and herbarium specimens) usually do not require special treatment for SEM. They are sufficiently dried in air. Seeds were transferred with a special tool to previously prepared special tables covered with double adhesive carbon tape. To minimize the static effect, the samples were sprayed with golden powder for 1-2 minutes through the JEOL JFC-1100E (JEOL Ltd., Tokyo, Japan) ion sputter coater.
Seeds can also be examined in the low vacuum mode without metal spraying. After that the table was placed in a microscope for SEM analysis. A general view of the seed surface and the observed features of the sculpture at high and low magnifications were photographed with a JEOL JSM6610 LV electron microscope (JEOL Ltd., USA). This microscope photographed nearly 1500 times or more in order to explore the microstructure of the seed's surface. Sometimes it was found useful to tilt the table; for example, to consider a weakly expressed sculpture. Also, on the cracked objects we can see the internal structure of the shells, which is also more convenient to do by tilting the table. The obtained data were stored on removable storage media. Morphological characters, including general shape, color and size of seeds, were studied under the stereomicroscope Leica EZ4D (USA).
Table 1
Deskriptor information about collected seed samples
Seed patterns Locality code Collection area Biotope Geographical coordinates Altitude, m
V. monantha AZE16D6 Shamakhi district, Shamakhi-Agsu highway roadside N 40°38'550 E 48°28'450 794
V. tenuifolia subsp. variabilis AZE17K19 Shamakhi region, Pirgulu village forest N 40°46'864 E48°36'168 1430
V. villosa subsp. villosa AZE17K20 Shamakhi region, Mirzandiya village meadow N 40°34737 E 48°43'648 584
V. varia AZE15Z2 Zagatala region, Yukhari Tala village, Parzivan area roadside N 46°35'29 E 41°34'227 358
V. elegans AZE16E4 Ordubad region, Tivi village foothills N 39°06'476 E 45°54702 1920
The most commonly accepted classification is the classification of Kupicha, so we accepted that classification in own work [23]. However, it should be noted that new phylogenetic results indicate that the genus Vicia, as traditionally understood, is not monophyletic [8; 9; 10; 31]. We used terminology adopted by some authors cited in our article [7; 10; 13; 16; 19; 20; 21; 28; 30; 32].
Results and discussion
During expeditions were collected seeds of taxa of Vicia belonging to subgenus Vicilla and section Cracca (according to the system of Kupicha): V. tenuifolia Roth subsp. variabilis (Freyn & Sint. ex Freyn) Dinsm., V. villosa subsp. villosa Roth, V. villosa Roth subsp. varia (Host) Corb. (V. dasycarpa auct.), V. monantha Retz. (V. cinerea M. Bieb.), V. elegans Guss.
The distribution of Vicia taxa collected from different research areas and descriptions of the collected seed samples are provided below:
Subgenus: Vicilla (Schur) Rouy
Sect.: Cracca S. F. Gray - Annual to perennial, often climbing. Stipules without a dark nectariferous spot. Leaflets few- many- paired, small to large. Peduncle usually much longer than the 1- many flowers. Flowers usually blue- lilac to purplish- red, rarely yellow. Calyx usually irregular and gibbous. Style compressed, equally pubescent all round near the apex. Legumes with coriaceous, glabrous or hairy valves, not torulose; seeds few to many.
Seed patterns: V tenuifolia Roth subsp. variabilis (Freyn & Sint. ex Freyn) Dinsm., V. villosa Roth subsp. villosa, V villosa Roth subsp. varia (Host) Corb. (V. dasycarpa auct.), V monantha Retz. (V. cinerea M. Bieb.), V. elegans Guss.
a j H C
« s e f
h I
r
m „ O
Fig. 1 General shape, hilum shape and seed sculpture some of examined taxa (SEM) a-c: V. tenuifolia subsp. variabilis; d-f: V. villosa subsp. villosa; g-i: V. villosa subsp. varia; j-1: V. monantha ; m-o: V. elegans (scale bar: a-c lmm, 500,50 \im; d-f 1mm, 200,50 \im; g-i 500,200,10 ¿un; j-11 mm, 500,50 ¿un; m-o 500,200,50 \im)
Micromorphological structure characteristics of seeds have been analyzed and important diagnostic elements have been identified that may be useful for systematics of controversial taxa. The majority of seed samples are spherical and oblong. The largest seeds are found in V. tenuifolia subsp. variabilis, and the smallest ones in V. villosa subsp. varia. Seeds are stained and striped. Hilum can be different of different patterns: long, loose or oval. In all seed samples, the seed coat is papillate. Papillas are large and small. Density, size and shape of papillas differ depending on the type of the papilla. The density of the papillae is estimated to be moderate to high.
Table 2
Testa texture of examined taxa
Signs // Seeds Seed shape Seed size (mm) Seed colour Densy of papillas Shape of papillas Texture Shape of hilum Hilum size (mm)
V. tenuifolia subsp. variabilis sferical-elliptic 4,168 X 2,760 light brown, with black spot medium small conical multirib papillose oblong-elliptic 2,115 X 0,424
V. villosa subsp. villosa spherical 3,240 X 3,048 black, with brown spot density conical multirib papillose oblong-elliptic 1,297 X 0,448
V. villosa subsp. varia round 2,821 x 2,771 black density conical, multirib, with waxy speckled papillose ovate 1,319 X 0,467
V. monantha spherical 3,903 X 3,711 light brown, with dark spot density small conical, multirib papillose oblong-elliptic 2,040 X 0,309
V. elegans ovate 3,947 X 3,083 black, marble density broad conical papillose oblong-elliptic 1,462 X 0,281
In this study, some characteristics of the seeds were used for the taxonomic delimitation and the identification of them was held according to these characteristics (Table 2). Ovate hilum was observed in V. villosa subsp. varia. Oblong-elliptic hilum is peculiar to V. tenuifolia subsp. variabilis, V. villosa subsp. villosa, V. monantha, and V. elegans. The papillae with medium density were observed in V. tenuifolia subsp. variabilis (Fig. 1. a-c). Dense papilla were observed in V. villosa subsp. villosa (Fig. 1. d-f), V. villosa subsp. varia (Fig.l. g-i), V. monantha and V. elegans (Fig. 1. j-1, Fig.l. m-o). The result showed that V. villosa subsp. varia had conical, multirib papillas with waxy specks. However, small conical multirib were observed in V. tenuifolia subsp. variabilis and V. monantha. Conical multirib was found in V. villosa subsp. villosa and broad conical in V. elegans.
General shape, colour and size of seeds were studied under a stereomicroscope (Fig.2).
Fig. 2 The general appearance of the seeds under stereomicroscope 1. V. tenuifolia subsp. variabilis; 2. V. villosa subsp. villosa; 3. V. villosa subsp. varia; 4. V. monantha; 5. V.
elegans
Conclusion
Micromorphological analysis of the seeds of important taxa of the genus Vicia L. was carried out to eliminate the gaps in seed identification in different Floras.
In result was detected that seeds of taxa of Vicia belonging to subgenus Vicilla and section Cracca had different micromorfological signs: Oblong-elliptic hilum is peculiar to V. tenuifolia subsp. variabilis, V. villosa subsp. villosa, V. monantha, and V. elegans (only ovate hilum in V. villosa subsp. varia). Dense papilla were observed in V. villosa subsp. villosa, V. villosa subsp. varia, V. monantha and V. elegans (only V. tenuifolia subsp. variabilis had papillae with medium density). V. villosa subsp. varia had conical, multirib papillas with waxy specks. Other taxa had simple (without waxy specks) conical papillas.
Acknowledgement
The authors are deeply grateful to the Institute of Geology and Geophysics, ANAS, Sh. Sh. Bayramova for assisting us to work with the SEM, also researchers G. Sh. Guliyeva and G. F. Allahverdiyeva for help in comparing photos.
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Статья поступила в редакцию 11.20.2020 г.
Асадова К.В., Аскеров А.М. Микроморфология семян в некоторых таксонах Vicia, принадлежащих к секции Cracca из Азербайджана // Bull, of the State Nikita Botan. Gard. - 2020. - № 134.-P. 17-23.
Впервые некоторые таксоны рода Вики (Vicia L.) из Азербайджана были подвергнуты морфологическому и сканирующему электронно-микроскопическому (SEM) анализу: V. elegans Guss., V. monantha Retz. (V. cinerea M. Bieb.), V. tenuifolia Roth subsp. variabilis (Freyn et Sint. ex Freyn) Dinsm., V. villosa Roth subsp. varia (Host) Corb. (V. dasycarpa auct.), V. villosa subsp. villosa Roth.
Образцы семян были собраны в 2015-2017 годах из разных регионов Азербайджана (Закатала, Ордубад и Шамахи). Собранные таксоны были сгруппированы в подвид Vicilla и в секцию Cracca. В образцах структура поверхности, форма, плотность, поверхность или воскообразный слой папилл (почки с поверхностью всасывания) были изучены методом сканирующей электронной микроскопии (СЭМ).
Морфологические признаки, общая форма, размер, цвет, структура поверхности семян и размеры семян были изучены под стереомикроскопом во время исследования.
Ключевые слова: семена, SEM, вика, бобовые, внутривидовые, сечение, вид
УДК 582.572.225:581.4
DOI: 10.36305/0513-1634-2020-134-23-28
БИОЛОГИЧЕСКИЕ ОСОБЕННОСТИ РЕДКОГО ВИДА ALLIUM GRANDE LIP SKY В БАШКИРСКОМ ПРЕДУРАЛЬЕ
Ленвера Ахнафовна Тухватуллина, Лариса Михайловна Абрамова
Южно-Уральский ботанический сад-институт - обособленное структурное подразделение Уфимского федерального исследовательского центра РАН 450080, Республика Башкортостан, г. Уфа, ул. Менделеева, д. 195/3 E-mail: abramova.lm@mail.ru
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