Fh molecular mutations are detected by a change in the nucleotide, and a biochemical mutation can be detected by alteration in a biochemica reaction. ‘the methods of detection of morphological mutants have been developed mainly with Drosophila, Four methods, viz., (1) CIB method, (2) Muller's 5 method, (3) attached X-chromosome method, and (4) curly Jobe plum method are in common use for detection of mutations in Drosophila, Abrief description of each method is presented below: i. CIB Method: This method was developed by Muller for detection of induced sex linked recessive lethal mutations in Drosophila male, In this technique, C represents a paracentric inversion in large part of X-chromosome which suppresses crossing over in the inverted portion. The 1 is a recessive lethal, Females with lethal gene can survive only in heterozygous condition. ‘The B stands for bar eye which acts as a marker and helps in identification of flies. The 1 and B are inherited together because C does not allow crossing over to occur between them. The males with CIB chromosome do not survive because of lethal effect. ‘The important steps of this method are as follows: (a) A cross is made between CIB female and mutagen treated male. In F, half of the males having normal X-chromosome will survive and those carrying CIB chromosome will die. Among the females, half have CIB chromosome and half normal chromosome (Fig. 14.2). From F,, females with CIB chromosome and male with normal chromosome are selected for further crossing, Scanned by CamScannerNormal Male iB Bar agen treated Female oleae Fist = 9) VY x coss og| (t COB bar Female MT Normal MT Female Second cross mutation Induced Fig, 14.2. Mullor’s CIB method for detection of lethal induced mutations in X-chromosome of Drosophila, Mt denotes mutagen treated X.chromsome. (b) Now a cross is made between CIB female and normal male. This time the CIB female has one CIB chromosome and one mutagen treated chromosome received from the male in earlier cross. This will produce two types of female viz., half with CIB chromosome and half with mutagen treated chromosome (with normal phenotype). Both the progeny will survive. In case of males, half with CIB will die and other half have mutagen treated chromosome. Scanned by CamScannerr If-a lethal mutation was induced in mutagen treated X-chromosome, the remaining half males will also dig, resulting in absence of male progeny in the above cross, ‘Absence of male progeny in F2 confirms the induction of sex linked recessive lethal mutation in the mutagen treated Drosophila male. ii. Muller 5 Method: This method was also developed by Muller to detect sex linked mutation in Drosophila. This method is an improved version of CIB method. This method differs from CIB method in two important aspects. First, this method utilizes apricot recessive gene in place of recessive lethal in CLB method. Second, the female is homozygous for bar apricot genes, where: method. it is heterozygous for IB genes in CIB In this method, the mutation is detected by the absence of wild males in Fz progeny. ‘This method consists of following important steps (Fig. 14.3). Scanned by CamScannerNotrmal Male Maier 5 bar Mate apricot teal Mulagan trate cy yc ur] First rots agar opr x al Ie bar apricot Progony apn Intermating of Fy Second cross 6 o | fe x dj [ B B Muller § ar apricot ‘mutation Induced) Fig. 14.3. Muller's 5 method of for detection of induced mutation in X-chromosome of Drosophila, Mt denotes mutagen treated X-chromosome. a. A homozygous bar apricot female is crossed with mutagen treated male. In Fy we get two types of progeny, viz., heterozygous bar females and bar apricot (Muller) males. b. These F, are inter-mated. This produces four types of individuals. Half of the females are homozygous bar apricot, and half are bar heterozygous. Among the males, half are bar apricot (Muller 5) and half should be normal, [fa lethal mutation is induced, the normal male will be absent in the progeny. . Attached X-Method: is method is used to detect sex linked visible mutations in Drosophila, In this method a female in which two X-chromosomes are united or attached together is used to study the mutation (Fig. 14.4). Therefore, this method is known as attached Scanned by CamScannerX-method. The attached X females (X: cross gives rise to super females (3 and YY. Y) are crossed to mutagen treated male, This X), attached female (XY), mutant male (XY) ‘The YY individuals die and super female also usually dies. The surviving male has received X-chromosome from mutagen treated male and Y chromesome from attached X-female. Since ¥ chromosome does not have corresponding allele of X= chromosome, even recessive mutation will express in such male which can be easily detected, Attached X-Female Normal Male (XY) (XX) Mutagen treated Cross ‘Super Female usually dies Progeny Visible recessive mutation Fig. 14.4, Attach X-method for detection of induced visible mutation in chromosome ‘of Drosophila. Mt. denotes mutagen treated X-chromosome. Curly Lobe- ‘This method is used for detection of mutation in autosomes. In this method curly refers to curly wings, lobe to lobed eye and phim to plum or brownish eye. All these three genes are recessive lethal, Curly (CY) andl lobed (L) genes are located in one chromosome and plum (Pm) in another but homologous chromosome. Crossing over between these chromosomes cannot occur due to presence of inversion, Moreover, homozygous individuals for CYL or Pm cannot survive because Scanned by CamScannerive. Thus, this system is also known as of lethal effect. Only heterozygotes: st balanced lethal system, This method consists of following steps (Fig. 14.5). Curly lebo Plum Normal Malo Female mutagen treated yest coca of i x owt] | \ + + Curly Lobe Female Curly Lobe Plum Female Second cross Curly Lobe Female Curly Lobe Male Third cross cy cy ‘Ts ye cy Fig. 14.5, ig. Detection of induced autosomal recessive Mutation in Drosophila. Scanned by CamScannera, Across is made between curly Iobe plum (CYL/Pm) female and mutagen treated male, This produces 50% progeny as curly lobe and 50% as plum. TABLE 144, Comparison of Different Methods of Detection of Mutation in Drosophila CIB Method Muller $ Method Atlachad X Method Curly lobe plum Method 1. Used to detect sex inked Used indetect sex linked Used to detect visible linked Jethal muta- recessive lethal muta- visible mutations. autosomes mutations. tions. tions, 1. Ulitises heterazygous Uses homozygous bar Uses attached X female, Used heterozygous curly CIB females. apricot females. lobe plum female, |. CID males dic due to No lethal effect in the YY individuals die to Homozygous individuals Fetal effect parents, ddve to lethal effect, for CYLIPM gene die. 1. Crossing over does not Crossing over does not Crossing over does not Crossing over docs not over occur. occur cxcut Involves two crosses Involves two crosses, Involves one eross only. Involves three crosses Mutation ‘is detected Mutation is detected by Males with visible Individuals with treated by the absence of the absence oF normal mutations are found. —autosomes show visible mates in the progeny males inthe progeny of ‘mutations. of second cross. secand crass. b. In the second generation cross is made between curly lobe female and curly lobe phim male, This will give rise to curly lobe plum, curly lobe and plum individuals in t £121 ratio and homozygous curly twill die due to lethal effect. From this progeny, curly lobe females and males are selected for further mating. c. In third generation, a cross is made between curly lobe female carrying one mutagen treated autosome and curly lobe male also carrying treated autosome. This results in production of 50% progeny as curly lobe, 25% homozygous curly lobe which die and 25% progeny homozygous for treated autosomes. ‘This will express as autosomal recessive mutation and constitute one third of the surviving progeny. A comparison of different methods of detection of mutation in Drosophila is given in Table 14.4. Detection of Mutations in Plants: As stated earlier, the techniques of cetection of induced mutations have been mostly developed on Drosophila, In plants, such techniques have not been developed properly. In plants, two methods are used for detection of mutations depending upon the visibility of mutations. ‘These methods are briefly described below: Scanned by CamScanner