**6. Characterization and utilization of the collection**

Detailed evaluation, characterization, and cataloguing of germplasm accessions in the collections are very important to the utility of the collections by the local and world cotton research community interested in using 'candidate' accessions for breeding purposes. As mentioned above, during the periodic renewal of seed of germplasm accessions to maintain the collection, accessions are evaluated and characterized for the major characteristics and traits useful for both biodiversity estimates and cotton improvement. Each collection has its independent programs and objectives toward this goal; therefore, there was a need to develop a unified germplasm evaluation methodology and descriptors [11; 12].

#### **6.1. Cotton germplasm descriptor and database**

As reported by Campbell et al. [3], all germplasm related activities and maintenance of the Uzbekistan cotton collections are funded by the Committee for Coordination of Science and Technology Development under the Cabinet of Ministry of Uzbekistan, MAWR, and ASUz where funds are given as competitive research and "a unique facility" maintenance grants. Moreover, each institution receives international grants for projects that utilize the germplasm

As highlighted in the recent reports by Campbell et al. [3] and Abdullaev et al. [8], all existing cotton germplasm resources, deposited and preserved in the country, are available for cotton researchers, breeders, and students who are conducting the cotton research. Written requests should be addressed to the directors of institutions, and should describe the purpose of the germplasm usage and specify the amount of seeds requested. Directors of institutions, with approval and on the responsibility of germplasm curators, grant the distribution of requested germplasm. If a specific germplasm with commercial interest is requested, a bi-lateral agree‐ ments are sought defining the condition of sharing that varies according institutions' internal regulations. All granted requests are registered in the germplasm exchange book of the

Previously, germplasm requested by foreign institutions were officially sent under a standard "research purposes only" Material Transfer Agreement (MTA) and accompanied by proper phytosanitary certificates upon approval of the National Coordinator of Plant Genetic Resources (PGR) of Uzbekistan and responsible offices at the MAWR. Presently, any genetic and biological material sharing is granted by approval of Committee for Coordination of Science and Technology Development under the Cabinet of Ministry of Uzbekistan with "a

Detailed evaluation, characterization, and cataloguing of germplasm accessions in the collections are very important to the utility of the collections by the local and world cotton research community interested in using 'candidate' accessions for breeding purposes. As mentioned above, during the periodic renewal of seed of germplasm accessions to maintain the collection, accessions are evaluated and characterized for the major characteristics and traits useful for both biodiversity estimates and cotton improvement. Each collection has its independent programs and objectives toward this goal; therefore, there was a need to

develop a unified germplasm evaluation methodology and descriptors [11; 12].

resources, and therefore, budget funding for germplasm related works [3].

**5. Sharing and exchange**

296 World Cotton Germplasm Resources

research purpose only" MTA.

**6. Characterization and utilization of the collection**

collection.

In UzRIPI, there is a computerized "CAC-DB" database for plant germplasm resources developed for Central Asian and Transcaucasian countries using Microsoft FoxPro [36; 37]. In this database, the principle information systems of the International Centre for Agricul‐ tural Research in the Dry Areas (ICARDA) and the N.I. Vavilov Research Institute of Plant Industry (VIR) is used. Major characteristics for the entire cotton collection of the UZRIPI as well as UzSRICBSP have been described in an electronic catalogue in Microsoft Excel (Dr. Fyzulla Abdullaev, germplasm specialist of the UzSRICBSP, personal communication).

Within the framwork of USDA-Uzbekistan cooperation programs, CGB scientists in collabo‐ ration with IG&PEB colleagues and cotton germplasm Unit staff worked on updating the cotton cataloguing system. Scientists suggested the use of a modified version of the "Descriptor for Cotton" which was originally developed and approved by IPGRI in 1985 (http://www/ ipgri/cgiar.org). After considering recommendations from more recent descriptors for plant germplasm resources (e.g., Descriptor for Groundnuts-1992; and Descriptor for Pista‐ chio-2002) and consulting with IG&PEB germplasm curator and leaders (Dr. Abdumavlyan Abdullaev and Dr. Sofiya Rizaeva), some modifications were introduced to the "Descriptor for Cotton-1985" of IPGRI. In the modified descriptors, two categories: "characterization and preliminary evaluation" and "further characterization and evaluation" were combined into one category "characterization and evaluation" because of the lack of preliminary information for accessions that were brought from abroad to the IG&PEB collection. New sub-categories, such as 1) cotton boll size, and 2) natural leaf defoliation properties were introduced; in "Susceptibility to biotic stresses" diseases and disease sources, specific to Uzbekistan, were introduced. At present, about 1,000 Upland germplasm accessions have been characterized for main agronomic, fibre quality properties and molecular diversity [12; 23; 24] using the modified "Descriptor for Cotton-1985" [Table 2; 8].

Further, ~1000 *G. hirsutum* cotton accessions were selected from IG&PEB cotton collection and evaluated for major morphological, agronomic and fibre quality traits in the Uzbekistan and Mexican growing environments in collaboration with Germplasm Unit of USDA-ARS at College Station, Texas, USA [12; 23; 24; 38]. Data from the evaluations was collected according to above mentioned descriptor. Consequently, pc-GRIN software package (used in creating US plant germplasm database) and Microsoft Access software packages were used to develop initial basic electronic database of IG&PEB collection. This initial database contains most of information about accessions and morphological trait data, including fibre quality and yield properties as well as molecular-genetic data for each accession (such as in microsatellite marker genotypes, molecular diversity, etc.). There is ongoing effort to expand this database through adding additional germplasm accessions from periodic germplasm seed renewal evaluations. There is a need to apply the same procedure for all existing collections and coordinate future joint efforts toward this goal.


**No Trait**

23 PETAL colour

24 PEATAL SPOT

25 POLLEN colour

26 PHOTO

29 BOLL SH

30 BOLL OPEN

35 SEED FUZZ

36 FUZZ colour

**Characteristic or short**

20 MONO Numeric Number of monopodia 21 SYMP Numeric Number of sympodia 22 STEAMNODE Numeric Number of total stem nodes

**Value Notes**

Petal colour

Pollen colour

Boll shape

Fuzz colour

Level of Boll opening

applied to cultivated plants only)

Fuzz grade (amount of fuzz on seeds)

Sensitivity to day/night length

Cotton Germplasm Collection of Uzbekistan http://dx.doi.org/10.5772/58590 299

<sup>9</sup> Petal spot

1

1 2 3

1 2 3

32 BOLL WEIGHT Boll weight (g) Numeric Average weight in grams of a 10 boll sample

1 2 3

<sup>34</sup> SEED WEIGHT Numeric Weight of 100 seeds in grams (could be

27 FLOWERING Numeric Days to 50% of plants flowering 28 OPENING Numeric Days to 50% of bolls opening

**description**

White Cream Light yellow Yellow Lavender Red bicolour

absent present

White Cream Yellow Purple

Round Oval Conical

Normal Intermediate Strom-proof

Fuzzy Sparse fuzz Naked seed

White Green Grey Brown (Tan)

31 BOLL NUM Numeric Number of boll

33 LOCULES NUM Numeric Locules per boll

37 LINT colour White 1 Lint colour

Not photoperiodic Slightly photoperiodic Photoperiodic Strictly photoperiodic


**No Trait**

3 ACCESSION NUMBER

298 World Cotton Germplasm Resources

7 HOMOGENIETY

8 GROW HABIT

9 ANTOCYAN

10 STEM HAIRINESS

12 LEAFSHP

18 BRANCHTYPE

<sup>5</sup> YEAR Year of acceptance to

**Characteristic or short**

2 ACCESSION NAME Name of line or cultivar Ex.: TM-1, 3-79

6 RENEWAL Last time of seeds renewal DD.MM.YYYY

Prostrate 3

Erect 7

Weak 0

Middle 1 Strong 2 Not became brown 3

Naked 1

Very weak 2 Weak 3 Middle 5 Strong 7 11 LEAF HAIRINES As above Hairiness of the leaves

Palmate(normal) 1

Digitate(okra) 3 Lanceolate(super okra) 4 13 LOBE NUMB Numeric Number of leaf lobe

0-5 cm 1 5-10 cm 2 10-15 cm 3 15-20 cm 4 19 HIEGHT (cm) Numeric Plant height in cm.

<sup>16</sup> SEEDTERM % Seed energy in thermostat (%)

Branching Type

Number of accession in

**Value Notes**

Not uniform 0 Homogeneity of cotton germplasm

Compact 5 Growth of habit (Bush shape)

YYYY

*Gossypium hirsutum spp. mexicanum*)

Colour of the plant in general

Hairiness of the plant stem

Numeric The energy required for seed germination in

first fruit brunch in cm

Distance of first sympodial fruit branch from

thermostat

<sup>1</sup> SCIENTIFIC NAME Latin name Name of genus, species, subspecies etc. (ex.:

collection catalogue Ex.: 1, 2, 3… 4 ORIGIN Place of sample origin Collection, author/scientist, country etc.

Uniform 1 population

Leaf shape Semi-digitate(semi-okra) <sup>2</sup>

14 EMERGENCE Numeric Days to emergence of cotton seedlings 15 SEEDENERGY % Numeric The energy required for seed germination

17 BRHGHT (HS) Numeric Height of the first fruit branch in cm.

**description**

collection


**6.2. Germplasm characterization and utilization**

Sanamyan, personal communication).

Cotton germplasm Units of each collection work extensively on characterization of their own germplasm resources within the framework of various projects. IG&PEB cotton germplasm Unit scientists, as mentioned above, investigate the evolution, taxonomy, phylogeny, hybrid‐ ization compatibility, usage and introgression of wild germplasm. On the basis of investiga‐ tions of the gene pools from IG&PEB collection, Abdullaev et al. [39] updated the Mauer taxonomy system for *Gossypium* L. and highlighted new species discovered at the end of XX century in their updated taxonomy, which was unknown for Mauer [40]. Researchers also have created hybridization compatibility schemes and identified phylogenetic relations between different cotton species [41]. Accordingly, the IG&PEB collection has developed a number of hybrids within and between different gene pools and cotton species. Cotton species and gene pools preserved in IG&PEB collection were studied for morphological and anatomical characteristics in detail and an "Atlas of *Gossypium* L. genus" has been published [41].

Cotton Germplasm Collection of Uzbekistan http://dx.doi.org/10.5772/58590 301

Being a breeding collection of cultivated cottons in Uzbekistan, the UzSRICBSP researchers mainly use their collection to study genetics of various agronomic traits and disease resistance aspects of germplasm accessions during the variety development process [7; 16]. The NU collection, in particular the genetic stocks and cytogenetic stocks has been characterized in detail for the genetic traits [17] and cytogenetic aberrations [19; 20; 42]. The unique cytogenetic collection preserved at the NU also is being characterized using molecular markers and identification of chromosome specificities of the monosomics is in progress (Dr. Marina

Cotton germplasm resources were the foundation to develop highly adapted, disease resistant and superior agronomic quality cotton cultivars for Uzbekistan that were highlighted in a number of past reports [2; 3; 6; 8; 10]. In particular, forecasting the benefit of utilization of wild cotton germplasm accessions in conventional breeding of cotton in early 1960s by germplasm leaders, Drs. F. Mauer and A. Abdullaev [3; 8; 43] boosted the new variety development in Uzbekistan that provided timely responses to cotton production problems in the country. Over the past several decades, Uzbekistan breeders have developed more than 200 new cotton cultivars highly adapted [7] to the different soil-climatic regions of Uzbekistan, with high resistance characteristics to the major cotton diseases and pests, having high-yield and better fibre qualities [6; 7; 8; 44]. It is noteworthy to highlight here that the major accomplishment of Uzbekistan cotton breeders for the past century of efforts was the development of very earlymaturing and productive cotton cultivars with 105-120 days of vegetation period, which made cotton growing suitable and profitable in the northern latitudes like Uzbekistan. This was only possible because of the existence and efficient exploitation of cotton germplasm resources that should be highlighted and valued as Uzbekistan's cotton breeders greatest commitment.

Further, as highlighted in recent reports [3; 8; 21] with the advances in cotton sciences and the development and application of modern molecular genetics tools and approaches for plant germplasm analysis [45], initial efforts by Uzbekistan scientist to characterize a selected global set of ~1000 *Gossypium hirsutum* L. accessions from Uzbekistan cotton germplasm at molecular level have occured. This global set represented 37 cotton growing countries and 8 breeding ecotypes as well as wild landrace stocks. Using this selected set of global Upland germplasm

**Table 2.** The descriptor developed for Uzbek cotton germplasm collection (Source: from Abdullaev et al., 2013)

#### **6.2. Germplasm characterization and utilization**

**No Trait**

300 World Cotton Germplasm Resources

**Characteristic or short**

38 fibre LENGHT Numeric fibre length in millimeter

41 UI% Numeric fibre maturity in %

47 CGRD Numeric colour grade of fibre 48 AREA Area of seed in % Numeric Seed covered area with fibre

> Resistant (≥76%-85%) 2 Moderate (≥51%-75%) 3 Susceptible (≥31%-50%) 4 Highly susceptible (1%-30%) 5

environmental conditions

Highly Resistant (≥86%

45 LINT INDEX Numeric The weight of lint from 100 seed in gram

Numeric

1

**Table 2.** The descriptor developed for Uzbek cotton germplasm collection (Source: from Abdullaev et al., 2013)

**Value Notes**

units

The fibre strength of a bundle of fibres measured on a Stelometre with the jaws holding the fibre bundle tightly apprised. Measured in grams force per tex.

The fitness of the sample taken from the ginned lint measured by the Micronaire and expressed in standard curvilinear micronaire

Hunter's B value, a measure of increasing yellowness of the cotton, taken with a Nickerson-Hunter colourimeter

A measure of the percentage of reflectance on a Nickerson-Hunter colourimeter. The higher the value the lighter the cotton.

The weight of lint ginned from sample of seed cotton expressed as percentage of the

The percentage of elongation at break of the

centre 1/8 each of the fibre bundle measured for T1 strength on the Stelometer

Percentage of plants Resistant to pests/ insects or phytopathogen (fungi, bacteria,

weight of seed cotton

virus etc.)

As above Resistance to salt, drought, low water, temperature etc.

**description**

Cream Light brown Brown Green

39 T0 fibre strength (g/tex) Numeric

40 MIC Micronaire Numeric

42 YELLOWNESS fibre yellowness Numeric

43 REFLECT Reflectance (RD) Numeric

44 LINT Lint (yield) percentage Numeric

elongation


<sup>46</sup> ELO The percentage of fibre

<sup>50</sup> ENVRES Special resistance to

49 DISEASE RES

Cotton germplasm Units of each collection work extensively on characterization of their own germplasm resources within the framework of various projects. IG&PEB cotton germplasm Unit scientists, as mentioned above, investigate the evolution, taxonomy, phylogeny, hybrid‐ ization compatibility, usage and introgression of wild germplasm. On the basis of investiga‐ tions of the gene pools from IG&PEB collection, Abdullaev et al. [39] updated the Mauer taxonomy system for *Gossypium* L. and highlighted new species discovered at the end of XX century in their updated taxonomy, which was unknown for Mauer [40]. Researchers also have created hybridization compatibility schemes and identified phylogenetic relations between different cotton species [41]. Accordingly, the IG&PEB collection has developed a number of hybrids within and between different gene pools and cotton species. Cotton species and gene pools preserved in IG&PEB collection were studied for morphological and anatomical characteristics in detail and an "Atlas of *Gossypium* L. genus" has been published [41].

Being a breeding collection of cultivated cottons in Uzbekistan, the UzSRICBSP researchers mainly use their collection to study genetics of various agronomic traits and disease resistance aspects of germplasm accessions during the variety development process [7; 16]. The NU collection, in particular the genetic stocks and cytogenetic stocks has been characterized in detail for the genetic traits [17] and cytogenetic aberrations [19; 20; 42]. The unique cytogenetic collection preserved at the NU also is being characterized using molecular markers and identification of chromosome specificities of the monosomics is in progress (Dr. Marina Sanamyan, personal communication).

Cotton germplasm resources were the foundation to develop highly adapted, disease resistant and superior agronomic quality cotton cultivars for Uzbekistan that were highlighted in a number of past reports [2; 3; 6; 8; 10]. In particular, forecasting the benefit of utilization of wild cotton germplasm accessions in conventional breeding of cotton in early 1960s by germplasm leaders, Drs. F. Mauer and A. Abdullaev [3; 8; 43] boosted the new variety development in Uzbekistan that provided timely responses to cotton production problems in the country. Over the past several decades, Uzbekistan breeders have developed more than 200 new cotton cultivars highly adapted [7] to the different soil-climatic regions of Uzbekistan, with high resistance characteristics to the major cotton diseases and pests, having high-yield and better fibre qualities [6; 7; 8; 44]. It is noteworthy to highlight here that the major accomplishment of Uzbekistan cotton breeders for the past century of efforts was the development of very earlymaturing and productive cotton cultivars with 105-120 days of vegetation period, which made cotton growing suitable and profitable in the northern latitudes like Uzbekistan. This was only possible because of the existence and efficient exploitation of cotton germplasm resources that should be highlighted and valued as Uzbekistan's cotton breeders greatest commitment.

Further, as highlighted in recent reports [3; 8; 21] with the advances in cotton sciences and the development and application of modern molecular genetics tools and approaches for plant germplasm analysis [45], initial efforts by Uzbekistan scientist to characterize a selected global set of ~1000 *Gossypium hirsutum* L. accessions from Uzbekistan cotton germplasm at molecular level have occured. This global set represented 37 cotton growing countries and 8 breeding ecotypes as well as wild landrace stocks. Using this selected set of global Upland germplasm resources, molecular genetic diversity, population diversification and strata, and the extent of linkage disequilibrium (LD) for the cotton genome were characterized [10; 11; 12; 23; 24].

better characterize the selected cotton germplasm resources, AM and NAM panels that require

Cotton Germplasm Collection of Uzbekistan http://dx.doi.org/10.5772/58590 303

Due to the importance of cotton production for the country and the historic expertise on cotton farming and production developed during the past century, Uzbekistan has prioritized and promoted the breeding programs, leading to a collection of one of the richest cotton germplasm resources in the world. Developed and maintained by several leading cotton research institu‐ tions in Uzbekistan, cotton germplasm collections preserve over 25,000 major cotton germ‐ plasm accessions of *Gossypium* species, wild, primitive, pre-domesticated landraces stocks, cultivar and breeding lines, mutants, genetic and cytogenetic stocks, and various types of within and between cotton gene pool hybrids. These resources were collected from the centres of origins of *Gossypium* species during scientific expeditions and through germplasm exchange with world germplasm centres, and developed by scientists and breeders of Uzbekistan for the past century. Consequently, these germplasm resources were the basis for the development of Uzbekistan's main commercial cultivars with early maturity and high productivity, making cotton profitable in the northernmost cotton growing country. This greatest achievement and commitment of cotton breeders timely responded and solved many past and current problems of cotton production as well as it will help to sustain the cotton production in Uzbekistan in

Although each cotton collection has its own specific goals and objectives for preserved materials, there is a huge need for conducting joint re-inventory for clarification of redundan‐ cies between some of collections. There is a need for building short-term and long term cold storage rooms for all collections available in the country to better preserve the collection for future generations and cotton production. Moreover, although initial efforts have been made, coordinated efforts are needed to create a unified electronic database to systematize the germplasm records for all collections that will help for better utilization of the accessions in

There are over 5,000 novel germplasm resources that were developed for past decade of collaborative efforts on molecular genetic characterization of cotton germplasm resources and establishment of modern MAS and GM programs. The modern cotton genomics and bioin‐ formatics programs in combination with conventional cotton breeding efforts will further enhance the characterization and utilization as well as efficient documentation and systemat‐

We thank the Uzbekistan government for continual funding support for cotton germplasm collections and research. We acknowledge the Office of International Research Programs

additional funding, coordinated efforts and international collaborations.

**7. Conclusion**

the future.

the breeding programs.

**Acknowledgements**

ization of the germplasm resources in Uzbekistan.

Efforts also identified several SSR markers associated with main fibre quality traits along with donor accessions, bearing "golden" QTLs useful for MAS programs [23; 24]. As a result, a successful MAS program has been established in Uzbekistan that facilitated to mobilize novel haplotypes of important fibre quality QTLs from donor accessions preserved in cotton germplasm collection to commercial cultivars. In that fibre trait-associated DNA markers were used as a tool to manipulate the transfer of QTLs during a genetic hybridization [38; 46; 47].

The MAS program, extensively utilized during past decade in Uzbekistan that involved several donor (germplasm accessions) and recipient (commercial cultivars) germplasm not only improved and developed novel MAS cultivars through mobilization of novel untapped loci but also created novel germplasm resources useful for cotton breeders and future cotton breeding [Table 1; 10; 38; 46; 47]. Several first generation MAS-derived cultivars were devel‐ oped and submitted for State Variety Testing Committee of Uzbekistan for future commerci‐ alization that are being field-trailed for improved agronomic and fibre qualities across the Uzbekistan regions. Moreover, as a ongoing effort, Uzbekistan researchers initiated a genepyramiding approach to combine major fibre quality traits into single genotype of several commercial Upland cotton cultivars of Uzbekistan using these effective molecular markers as a breeding tool and donor genotypes from cotton germplasm collections [38; 46; 47].

As a continuation of efforts to characterize cotton germplasm resources at the molecular level, about 300 *G. barbadense* (so called extra long staple fibre Pima cotton) accessions with wide geographic origin covering 17 countries worldwide were recently genotyped with SSR markers to estimate molecular diversity, phylogeny, population structure, and linkage disequilibrium level. These Pima cotton accessions were exchanged with USDA-ARS at Shafter, California, evaluated for major agronomic, morphological and fibre quality traits in the Uzbekistan or/and CA growing environments, and used for molecular mapping of fibre quality traits useful for MAS programs [8; Abdullaev et al., 2014 unpublished]

Use of cotton germplasm resources and their characterization at the molecular level further helped us to select the most diverse *G. hirsutum* accessions to create nested association mapping (NAM) panel. The creation of NAM populations is efficient approach to enhance power [48] of association mapping in cotton [10; 49]. Currently, a panel of 4000 F2:3 generation NAM population individuals has been developed from genetic crosses between 20 most diverse *G. hirsutum* accessions and Uzbek cultivar Namangan-77, a genetic standard for *G. hirsutum.* NAM population individuals are also being genotyped with a large number of SSR markers and evaluated for major agronomic characteristics in different environments of Uzbekistan to conduct fine association mapping studies [22; Abdurakhmonov et al., 2014, unpublished].

Currently, genotyping of additional cotton germplasm accessions from major collections using widely available SSR markers is in progress that will help to characterize and organize the entire cotton germplasm resources of Uzbekistan. In perspective, we also aim to apply novel molecular tools such as SNP platforms and genotyping-by-sequencing (GBS) technologies to better characterize the selected cotton germplasm resources, AM and NAM panels that require additional funding, coordinated efforts and international collaborations.
