Features of Chromosome Introgression from Gossypium barbadense L. into G. hirsutum L. during the Development of Alien Substitution Lines

Abstract

The creation of G. barbadense L./G. hirsutum L. chromosome-substitution lines is an important method to transfer agronomically valuable traits from G. barbadense into G. hirsutum. In this study, 30 monosomic lines of G. hirsutum from the Cytogenetic Collection of Uzbekistan, created in the genotypic background of line L-458, were used in crosses with the G. barbadense line Pima 3-79 to create substitution lines. In the course of this work, new monosomic lines were identified for chromosome 12 and monotelodisome 6 of the A_tsubgenome and for chromosomes 17, 21, and 22 of the D_tsubgenome using chromosome-specific SSR markers and a well-defined tester set of cotton translocation lines (USA). Compared to those in the F₁ hybrids, a strong decrease in the crossing and setting rates was found in the BC₁F₁ backcross lines, with the substitution of chromosomes 2, 4, 6, 7, and 12 of the A_tsubgenome and 17, 18, 21, and 22 of the D_tsubgenome. The F₁ and BC₁F₁ offspring from interspecific crosses differed in their transmission of univalents. Despite the regular pairing of chromosomes and the high meiotic index, interspecific aneuploid hybrids were characterized by a decrease in pollen fertility, which may indicate hidden structural variability in these genomes that did not affect meiotic division. The identification of chromosomes using chromosome-specific SSR markers in the early stages of plant development has greatly accelerated the detection of monosomic plants. The analysis of morphobiological traits revealed that monosomic F₁ hybrids were more similar to the donor line, while BC₁F₁ hybrids were more similar to the recurrent parent but also showed previously undetected traits.

Keywords: G. barbadense; G. hirsutum; chromosome substitution lines; cotton; molecular markers; monosomic lines; translocation.

Figure 1

Development of cotton chromosome-substitution lines. The scheme shows the different steps for obtaining cotton chromosome-substitution lines, including cytogenetic analysis and the use of molecular markers (SSRs).

Figure 2

Crossing of aneuploid G. hirsutum L. lines with the G. barbadense L. line Pima 3-79.

Figure 3

Setting of F₀ hybrid seeds obtained from crosses of aneuploid G. hirsutum L. lines with the G. barbadense L. line Pima 3-79.

Figure 4

Germination of F₀ hybrid seeds obtained from crosses of aneuploid G. hirsutum L. lines with the G. barbadense L. line Pima 3-79.

Figure 5

Twenty-five normal bivalents and one heteromorphic bivalent obtained at Metaphase I of F₁ hybrids from a cross between a monotelodisome line Mo21 × Pima 3-79 (100₁) for a chromosome 11 arm with the line Pima 3-79. (Heteromorphic bivalent arrowed.) Scale bar = 10 µm.

Figure 6

Metaphase I in F₁ hybrids obtained from crosses of monosomic lines with the line Pima 3-79: (a) (Mo16 × Pima 3-79) (25^II + 1^I) (hybrid 98₂) with monosomy for chromosome 2; (b) (Mo34 × Pima 3-79) (25^II + 1^I) (hybrid 2₈) and (c) (Mo95 × Pima 3-79) (25^II + 1^I) (hybrid 108₃) with monosomy for chromosome 6; (d) monosomy for chromosome 12 (Mo94 × Pima 3-79) (25^II + 1^I) (8₁₃). (Univalents arrowed). Scale bar = 10 µm.

Figure 7

Metaphase I in meiosis of F₁ hybrids obtained from crosses of monosomic lines with the line Pima 3-79: (a) (Mo70 × Pima 3-79) (25^II +1^I) (hybrid 774₆) and (b) (Mo89 × Pima 3-79) (25^II + 1^I) (hybrid 515₇) with monosomy for chromosome 4; (c) monosomy for chromosome 17 (Mo56 × Pima 3-79) (25^II + 1^I) (4₁₃); and (d) monosomy for chromosome 18 (Mo48 × Pima 3-79) (25^II + 1^I) (529₁₆). (Univalents arrowed.) Scale bar = 100 µm.

Figure 8

Telophase II in F₁ hybrids: (a) Two tetrads without cytokinesis (Mo16 × Pima 3-79) (98₂); (b) anomalous triad (Mo60 × Pima 3-79) (694₅); (c) dyad with two micronuclei; and (d) tetrad with three micronuclei. Scale bar = 40 µm.

Figure 9

Telophase II in F₁(Mo67 × Pima 3-79) (308₁) hybrid: (a) normal tetrad; (b) two monads and two tetrads; (c) anomalous tetrad; (d) tetrad with a micronucleus; and (e) heptad. Scale bar = 40 µm.

Figure 10

Fertile (coloured) and sterile (uncoloured) pollen grains from a hybrid; (a) F₁ (Mo38 × Pima 3-79) (690₁₁) and (b,c) F₁(Mo89 × Pima 3-79) (515₇) for chromosome 4. Scale bar = 40 µm.

Figure 11

Chromosome identification of monosomic interspecific G. hirsutum × G. barbadense F₁ hybrids. SSR primer pair specific for chromosome 12 of the A_tsubgenome: BNL1707.

Figure 12

Chromosome identification of monosomic interspecific G. hirsutum × G. barbadense F₁ hybrids. SSR primer pair specific for chromosome 17 of the D_tsubgenome: (a) TMB0874; (b) BNL2496.

Figure 13

Chromosome identification of monosomic interspecific G. hirsutum × G. barbadense F₁ hybrids. SSR primer pair specific for chromosome 21 of the D_tsubgenome: BNL1705.

Figure 14

“Critical configurations” of the chromosomes at Metaphase I: 24^II + 1^III from cotton monosomic F₁ plants from the cross Mo42 × TT10L-21L. (Arrows point to the trivalents.) Scale bar = 100 µm.

Figure 15

Features of the monosomic cotton lines for chromosome 12: (a) bush, (b) configurations of the chromosomes (25^II + 1^I), (c) leaf, (d) flower, (e) petal, (f) bracts, (g) staminate column, (h) and (i) green bolls, (j) boll with peduncle, and (k) open boll.

Figure 16

Features of the monosomic cotton lines for chromosome 17: (a) bush, (b) configurations of the chromosomes (25^II + 1^I), (c) leaf, (d) flower, (e) petal, (f) bracts, (g) staminate column, (h) and (i) green bolls, (j) boll with peduncle, and (k) open boll.

Figure 17

Features of the monosomic cotton line for chromosome 21: (a) bush, (b) configurations of the chromosomes (25^II +1^I), (c) leaf, (d) flower, (e) petal, (f) bracts, (g) staminate column, (h) and (i) green bolls, (j) boll with peduncle, and (k) open boll.

Figure 18

Features of the monosomic cotton line for chromosome 22: (a) bush, (b) configurations of the chromosomes (25^II +1^I), (c) leaf, (d) flower, (e) petal, (f) bracts, (g) bract, (h) staminate column, (i) green bolls, (j) boll with peduncle, and (k) open boll.

Figure 19

Features of the monotelodisomic cotton lines for chromosome 6: (a) bush, (b) configurations of the chromosomes (25^II + Ii), (c) leaf, (d) flower, (e) petal, (f) bracts, (g) staminate column, (h) and (i) green bolls, (j) boll with peduncle, and (k) open boll.

Figure 20

Crossing of aneuploid lines of cotton G. hirsutum L. with the interspecific aneuploidy hybrid F₁ (Mo × Pima 3-79 or Telo × Pima 3-79).

Figure 21

Setting of BC₁F₁ hybrid seeds obtained from crosses of aneuploid lines of cotton G. hirsutum L. with interspecific aneuploid F₁ hybrids (Mo × Pima 3-79 or Telo × Pima 3-79).

Figure 22

Germination of BC₁F₁ hybrid seeds obtained from crosses of aneuploid lines of cotton G. hirsutum L. with interspecific aneuploid F₁ hybrids (Mo × Pima 3-79 or Telo × Pima 3-79).

Figure 23

Metaphase I in BC₁F₁ hybrids obtained from crosses of monosomic lines with interspecific monosomic F₁ (Mo × Pima 3-79) hybrids: (a) substitution of chromosome 6 (Mo92 × F₁539₅) (1040₂) (25^II + 1^I); (b) substitution of chromosome 12 (Mo94 × F₁8₃) (299₁) (25^II + 1^I); (c) substitution of chromosome 4 BC₁F₁ (Mo58 × F₁530₃) (115₁); (d) BC₁F₁ (Mo60 × F₁694₅) (117₅); and (e) BC₁F₁ (Mo75 × F₁104₂) (298₃) (25^II + 1^I). (Univalent arrowed). Scale bar = 10 µm.

Figure 24

Metaphase I in BC₁F₁ hybrids obtained from crosses of monosomic lines with interspecific monosomic F₁ (Mo × Pima 3-79) hybrids: (a) substitution of chromosome 7 (Mo27 × F₁687₄) (111₅) (25^II + 1^I); (b) substitution of chromosome 18 (Mo48 × F₁529₁₆) (114₁) (25^II +1^I); (c) substitution of chromosome 22 (Mo17 × F₁685₇) (110₁) (25^II + 1^I). (Univalent arrowed.) Scale bar = 10 µm.

Figure 25

Telophase II in a monosomic BC₁F₁ (Mo48 × F₁529₁₆) hybrid (114₁): (a) dyad; (b) pentad; (c) octad. Scale bar = 40 µm.

Figure 26

Fertile (coloured) and sterile (uncoloured) pollen grains in a aneuploid hybrid BC₁F₁ obtained from a cross between a aneuploid line and a aneuploid hybrid F₁ (Mo × Pima 3-79 or Telo × Pima 3-79): (a,b) BC₁F₁ (Mo60 × F₁694₅) (117₄); (c) BC₁F₁ (Mo34 × F₁688₉) (293₃); and (d) BC₁F₁ (Telo12 × F₁542₈) (561₁₅).

Figure 27

Chromosome identification of monosomic BC₁F₁ (Mo16 × F₁98₆) hybrids. SSR primer pairs specific for chromosome 2 of the A_tsubgenome JESPR179.

Figure 28

Chromosome identification of monosomic BC₁F₁ (Mo34 × F₁688₉) hybrids. SSR primer pairs specific for chromosome 6 of the A_tsubgenome Gh082.

Figure 29

Plants of the parental lines L-458 (a) and Pima 3-79 (b) disomic F₁ hybrid L-458 × Pima 3-79 (c) (Mo × Pima 3-79) and F₁BC₁ obtained from crosses of the recurrent parent; (d) monosomic line for chromosome 2, Mo16; (e) F₁(Mo16 × Pima 3-79) (98₂); and (f) substitution of chromosome 2 BC₁F₁(Mo16 × F₁98₂) (923₇).

Figure 30

Plants of monosomic line G. hirsutum for chromosome 4, Mo59 (a); F₁Mo59 × Pima 3-79 (531₂) (b); and BC₁F₁ (Mo59 × F₁531₂) (1041₄) with substitution for chromosome 4 G. barbadense (c).

Figure 31

Plants of monosomic line G. hirsutum for chromosome 6, Mo34 (a); F₁Mo34 × Pima 3-79 (688₉) (b); and BC₁F₁ (Mo34 × F₁688₉) (293₃) with substitution for chromosome 6 G. barbadense (c).

Figure 32

Plants of monosomic line G. hirsutum for chromosome 7, Mo27 (a); F₁Mo27 × Pima 3-79 (687₄) (b); and BC₁F₁ (Mo27 × F₁687₄) (111₂) with substitution for chromosome 7 G. barbadense (c).

Figure 33

Plants of monosomic line G. hirsutum for chromosome 12, Mo94 (a); F₁Mo94 × Pima 3-79 (8₁) (b); and BC₁F₁ (Mo94 × F₁8₁) (299₁) with substitution for chromosome 12 G. barbadense (c).

Figure 34

Plants of monosomic line G. hirsutum for chromosome 17, Mo56 (a); F₁Mo56 × Pima 3-79 (4₁₇) (b); and BC₁F₁ (Mo56 × F₁4₁₇) (513₈) with substitution for chromosome 17 G. barbadense (c).

Figure 35

Plants of monosomic line G. hirsutum for chromosome 18, Mo48 (a); F₁Mo48 × Pima 3-79 (529₁₆) (b); and BC₁F₁ (Mo48 × F₁529₁₆) (114₂₀) with substitution for chromosome 18 G. barbadense (c).

Figure 36

Plants of monosomic line G. hirsutum for chromosome 21, Mo42 (a); F₁ Mo42 × Pima 3-79 (528₁) (b); and BC₁F₁ (Mo42 × F₁528₁) (782₁) with substitution for chromosome 21 G. barbadense (c).

Figure 37

Plants of monosomic line G. hirsutum for chromosome 22, Mo17 (a); F₁ Mo17 × Pima 3-79 (685₇) (b); and BC₁F₁ (Mo17 × F₁685₇) (110₁) with substitution for chromosome 22 G. barbadense (c).

Figure 38

Plants of the parental line and monotelodisome F₁ and F₁BC₁ cotton hybrids obtained from crosses of the recurrent parent with monotelodisome F₁ hybrids (Telo12 × Pima 3-79): (a) monotelodisome line Telo12, chromosome 6; (b) F₁ Telo12 × Pima 3-79 (542₁₆); and (c) BC₁F₁ (Telo12 × F₁542₁₆) (561₁₅) with substitution of an arm of chromosome 6.

Figure 39

Plants of the parental line and monotelodisomic F₁ and F₁BC₁ hybrids obtained from crosses of the recurrent parent with monotelodisome F₁ hybrids (Telo × Pima 3-79): (a) monotelodisome line Mo21, chromosome 11; (b) F₁Mo21 × Pima 3-79 (102₁); and (c) BC₁F₁(Mo21 × F₁102₁) (292₁) with substitution of an arm of chromosome 11.